3-(5-hydroxy-1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives and uses thereof

ABSTRACT

The present disclosure provides a compound of Formula (I′):or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein Rx, X1, X2, and R1 are as defined herein, and methods of making and using same.

RELATED APPLICATIONS

This application claims the benefit of and priority to U.S. Provisional application Nos. 62/695,920, filed Jul. 10, 2018, and 62/835,543, filed Apr. 18, 2019, the entire contents of each of which are incorporated herein by reference in their entireties.

FIELD OF THE DISCLOSURE

The present disclosure relates to 3-(5-hydroxy-1-oxoisoindolin-2-yl)piperidine-2,6-dione compounds and compositions and their use for the treatment of IKAROS Family Zinc Finger 2 (IKZF2)-dependent diseases or disorders or where reduction of IKZF2 or IKZF4 protein levels can ameliorate a disease or disorder.

BACKGROUND OF THE DISCLOSURE

IKAROS Family Zinc Finger 2 (IKZF2) (also known as Helios) is one of the five members of the Ikaros family of transcription factors found in mammals. IKZF2 contains four zinc finger domains near the N-terminus which are involved in DNA binding and two zinc finger domains at the C-terminus which are involved in protein dimerization. IKZF2 is about 50% identical with Ikaros family members, Ikaros (IKZF1), Aiolos (IKZF3), and Eos (IKZF4) with highest homology in the zinc finger regions (80%+ identity). These four Ikaros family transcription factors bind to the same DNA consensus site and can heterodimerize with each other when co-expressed in cells. The fifth Ikaros family protein, Pegasus (IKZF5), is only 25% identical to IKZF2, binds a different DNA site than other Ikaros family members and does not readily heterodimerize with the other Ikaros family proteins. IKZF2, IKZF1 and IKZF3 are expressed mainly in hematopoietic cells while IKZF4 and IKZF5 are expressed in a wide variety of tissues. (John, L. B., et al., (2011), Mol. Immunol. 48:1272-1278; Perdomo, J., et al., (2000), J. Biol. Chem. 275:38347-38354.)

IKZF2 is believed to have an important role in the function and stability of regulatory T cells (Tregs). IKZF2 is highly expressed at the mRNA and protein level by regulatory T-cell populations. Knockdown of IKZF2 by siRNA has been shown to result in downregulation of FoxP3 and to impair the ability of isolated human CD4+ CD25+ Tregs to block T-cell activation in vitro. Moreover, overexpression of IKZF2 in isolated murine Tregs has been shown to increase expression of Treg related markers such as CD103 and GITR and the IKZF2 overexpressing cells showed increased suppression of responder T-cells. IKZF2 has also been found to bind the promoter of FoxP3, the defining transcription factor of the regulatory T-cell lineage, and to affect FoxP3 expression.

Knockout of IKZF2 within FoxP3-expressing Tregs in mice has been shown to cause activated Tregs to lose their inhibitory properties, to express T-effector cytokines, and to take on T-effector functions. IKZF2 knockout mutant mice develop autoimmune disease by 6-8 months of age, with increased numbers of activated CD4 and CD8 T cells, follicular helper T cells, and germinal center B cells. This observed effect is believed to be cell intrinsic, as Rag2−/− mice given bone marrow from IKZF2 knockout mice, but not bone marrow from IKZF2+/+ develop autoimmune disease. Direct evidence that IKZF2 affects regulatory T-cell function has been shown in the analysis of mice in which IKZF2 was deleted only in FoxP3 expressing cells (FoxP3-YFP-Cre Heliosfl/fl). The results showed that the mice also develop autoimmune disease with similar features as observed in the whole animal IKZF2 knockout. Moreover, pathway analysis of a CHIP-SEQ experiment has also suggested that IKZF2 is affecting expression of genes in the STAT5/IL-2Ra pathway in regulatory T-cells. This effect of IKZF2 loss was shown to be more apparent after an immune challenge (viral infection or injection with sheep's blood), and it was noted that after immune stimulation, the IKZF2 negative regulatory T cells began to take on features of effector T cells. (Getnet, D., et al., Mol. Immunol. (2010), 47:1595-1600; Bin Dhuban, K., et al., (2015), J. Immunol. 194:3687-96; Kim, H-J., et al., (2015), Science 350:334-339; Nakawaga, H., et al., (2016) PNAS, 113: 6248-6253)

Overexpression of Ikaros isoforms which lack the DNA binding regions have been shown to be associated with multiple human haematological malignancies. Recently, mutations in the IKZF2 gene, which lead to abnormal splicing variants, have been identified in adult T-cell leukemias and low hypodiploid acute lymphoblastic leukemia. It has been proposed that these isoforms, which are capable of dimerization, have a dominant negative effect on Ikaros family transcription factors which primes the development of lymphomas. IKZF2 knockout mutants that survive into adulthood do not develop lymphomas, supporting this hypothesis (Asanuma, S., et al., (2013), Cancer Sci. 104:1097-1106; Zhang, Z., et al., (2007), Blood 109:2190-2197; Kataoka, D., et al., (2015), Nature Genetics 47:1304-1315.)

Currently, anti-CTLA4 antibodies are used in the clinic to target Tregs in tumors. However, targeting CTLA4 often causes systemic activation of T-effector cells, resulting in excessive toxicity and limiting therapeutic utility. Up to ¾ of patients treated with a combination of anti-PD1 and anti-CTLA4 have reported grade 3 or higher adverse events (National Cancer Institute, Division of Cancer Treatment & diagnosis, Common Terminology for Adverse Events (CTCAE), https://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm). Thus, a strong need exists to provide compounds that target Tregs in tumors without causing systemic activation of T-effector cells.

An IKZF2-specific degrader has the potential to focus the enhanced immune response to areas within or near tumors providing a potentially more tolerable and less toxic therapeutic agent for the treatment of cancer.

SUMMARY OF THE DISCLOSURE

The compounds of the disclosure have use as therapeutic agents, particularly for cancers and related diseases. In one aspect, the compounds of the disclosure have IKZF2 degrader activity, preferably having such activity at or below the 50 μM level, and more preferably having such activity at or below the 10 μM level. In another aspect, the compounds of the disclosure have degrader activity for IKZF2 that is selective over one or more of IKZF1, IKZF3, IKZF4, and/or IKZF5. In another aspect, the compounds of the disclosure have degrader activity for both IKZF2 and IKZF4. The compounds of the disclosure have usefulness in treating cancer and other diseases for which such degrader activity would be beneficial for the patient. For example, while not intending to be bound by any theory, the inventors believe that reducing levels of IKZF2 in Tregs in a tumor may allow the patient immune system to more effectively attack the disease. In summary, the present disclosure provides novel IKZF2 degraders useful for the treatment of cancer and other diseases.

A first aspect of the present disclosure relates to compounds of Formula (I′):

wherein:

X₁ and X₂ are each independently H, (C₁-C₄)alkyl, (C₁-C₆)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₆)haloalkoxy, (C₃-C₇)cycloalkyl, halogen, —CN, —OH, or —NH₂;

R_(x) is H or D;

R₁ is

each R₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —CN, —OH, or —NH₂; or

two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₃ is (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —OH, or —NH₂;

R₄ is —OR₅ or —NR₆R_(6′);

R₅ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂; or

R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 8-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₉;

each R₈ is independently at each occurrence halogen, (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, —CN, —OH, —NR₁₃R₁₄, —NH₂, —O(C₃-C₇)cycloalkyl, —O-4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, —O(C₆-C₁₀)aryl, or —O-5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkoxy is optionally substituted with one to three R₁₀ and the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with one to three R₁₁; or

two R₈ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅; or two R₈ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S; or two R₈ together with the same atom to which they are attached form a ═(O);

each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, (C₃-C₆)cycloalkyl, —OH, —CN, —NH₂, or —NR₁₃R₁₄; or

two R₉ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one or more substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂; or two R₉ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

each R₁₀ is independently at each occurrence selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

each R₁₁ is independently at each occurrence selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, and —NH₂;

each R₁₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, or —NH₂;

two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S;

R₁₃ and R₁₄ are each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S;

m and m1 are each independently 0, 1 or 2;

n1 is 0, 1, 2, or 3; and

each s and n is independently 1, 2, or 3, wherein s+n is ≤4;

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In one embodiment, the present disclosure relates to compounds of Formula (I′) having the structure of Formula (I):

wherein:

R_(x) is H or D;

R₁ is

each R₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —CN, —OH, or —NH₂; or

two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₃ is (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —OH, or —NH₂;

R₄ is —OR₅ or —NR₆R_(6′);

R₅ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇; or

R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₉;

each R₈ is independently at each occurrence halogen, (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, —CN, —OH, or —NH₂, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S; or

two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S;

each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, or —NH₂; or

two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one or more substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

m and m1 are each independently 0, 1 or 2;

n1 is 0, 1, 2, or 3; and

each s and n is independently 1, 2, or 3, wherein s+n is ≤4;

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another aspect, the present disclosure relates to compounds of Formula (I′) or Formula (I′) or Formula (I), wherein:

R_(x) is H or D;

R₁ is

each R₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —CN, —OH, or —NH₂; or

two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₃ is (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —OH, or —NH₂;

R₄ is —OR₅ or —NR₆R_(6′);

R₅ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇; or

R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₉;

each R₈ is independently at each occurrence halogen, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —CN, —OH, or —NH₂; or

two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S;

each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, or —NH₂; or

two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one or more substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

m and m1 are each independently 0, 1 or 2;

n1 is 0, 1, 2, or 3; and

each s and n is independently 1, 2, or 3, wherein s+n is ≤4;

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In one aspect of the disclosure, the hydrogens in the compound of Formula (I′) or Formula (I′) are present in their normal isotopic abundances. In a preferred aspect of the disclosure, the hydrogens are isotopically enriched in deuterium (D), and in a particularly preferred aspect of the invention the hydrogen at position R_(x) is enriched in D, as discussed in more detail concerning isotopes and isotopic enrichment below.

Another aspect of the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition is useful in the treatment of IKZF2-dependent diseases or disorders. The pharmaceutical composition may further comprise at least one additional pharmaceutical agent.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient for use in the treatment of an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder. The pharmaceutical composition is useful in the treatment of IKZF2-dependent diseases or disorders. The pharmaceutical composition may further comprise at least one additional pharmaceutical agent.

Another aspect of the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition is useful in the treatment of diseases or disorders affected by the reduction of IKZF2 protein levels. The pharmaceutical composition may further comprise at least one additional pharmaceutical agent.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient for use in the treatment of a disease or disorder affected by the reduction of IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the disease or disorder. The pharmaceutical composition may further comprise at least one additional pharmaceutical agent.

In another aspect, the present disclosure relates to a method of degrading IKZF2. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a method of modulating IKZF2 protein levels. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a method of reducing IKZF2 protein levels. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a method of decreasing IKZF2 protein levels. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a method of reducing the proliferation of a cell. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof and reducing IKZF2 protein levels.

In another aspect, the present disclosure relates to a method of reducing IKZF2 protein levels. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a method of treating cancer. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In one embodiment, the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In another embodiment, the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In yet another embodiment, the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC). In another embodiment, the cancer is a cancer for which the immune response is deficient or an immunogenic cancer.

Another aspect of the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient for use in the treatment of an IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient in the manufacture of a medicament for treating of an IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a method for treating an IKZF2-dependent disease or disorder comprising the step of administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient.

In another aspect, the present disclosure relates to a method for treating an IKZF2-dependent disease or disorder comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating an IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a method for treating a disease or disorder that is affected by the modulation of IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a method for treating a disease or disorder that is affected by a decrease in IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a method for treating a disease or disorder that is affected by the reduction of IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by the modulation of IKZF2 protein levels.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by the reduction of IKZF2 protein levels.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by a decrease in IKZF2 protein levels.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by the modulation of IKZF2 protein levels.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by the reduction of IKZF2 protein levels.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by a decrease in IKZF2 protein levels.

In another aspect, the present disclosure relates to a method of treating cancer comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the cancer is a cancer for which the immune response is deficient or an immunogenic cancer.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease or disorder associated with the modulation of IKZF2 protein levels. In one embodiment, the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease or disorder associated with the modulation of IKZF2 protein levels. In one embodiment, the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease or disorder associated with the reduction of IKZF2 protein levels. In one embodiment, the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease or disorder associated with the reduction of IKZF2 protein levels. In one embodiment, the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease or disorder associated with a decrease in IKZF2 protein levels. In one embodiment, the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease or disorder associated with a decrease in IKZF2 protein levels. In one embodiment, the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

Another aspect of the present disclosure relates to a method of treating cancer comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of cancer, wherein the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating cancer, wherein the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to a method of treating a disease or disorder that is affected by the modulation of IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

Another aspect of the present disclosure relates to a method of treating a disease or disorder that is affected by the reduction of IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to a method of treating a disease or disorder that is affected by a decrease of IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by the modulation of IKZF2 protein levels wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by the reduction of IKZF2 protein levels wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by a decrease of IKZF2 protein levels, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by the modulation of IKZF2 levels, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by the reduction of IKZF2 protein levels, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by a decrease in IKZF2 protein levels, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma. In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC).

In another aspect, the present disclosure relates to a method of treating a disease or disorder that is affected by the modulation of IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

Another aspect of the present disclosure relates to a method of treating a disease or disorder that is affected by the reduction of IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

In another aspect, the present disclosure relates to a method of treating a disease or disorder that is affected by a decrease in IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by the modulation of IKZF2 protein levels, wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by the reduction of IKZF2 protein levels, wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by a decrease in IKZF2 protein levels, wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by the modulation of IKZF2 protein levels wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by the reduction of IKZF2 protein levels wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by a decrease in IKZF2 protein levels wherein the disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

In another aspect, the present disclosure relates to a method of treating cancer comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the cancer is a cancer for which the immune response is deficient or an immunogenic cancer.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a cancer for which the immune response is deficient or an immunogenic cancer.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a cancer for which the immune response is deficient or an immunogenic cancer.

Another aspect of the present disclosure relates to a method of treating an IKZF2-dependent disease or disorder by modulating IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to a method of treating an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a method of treating an IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the decrease in IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a method of treating an IKZF2-dependent disease or disorder by modulating IKZF2 protein levels comprising the step of administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to a method of treating an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels comprising the step of administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a method of treating an IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels comprising the step of administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein the decrease in IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by modulating IKZF2 protein levels wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by modulating IKZF2 protein levels wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels wherein the decrease in IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels wherein the decrease in IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of an IKZF2-dependent disease or disorder by modulating IKZF2 protein levels wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of an IKZF2-dependent disease or disorder by modulating IKZF2 protein levels wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of an IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels wherein the decrease in IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of an IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels wherein the decrease in IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a method of treating a disease or disorder by reducing IKZF2 protein levels comprising the step of administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein reduction of IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to a method of treating a disease or disorder by reducing IKZF2 protein levels comprising the step of administering to a subject in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein reduction of IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to a method of treating a disease or disorder by modulating IKZF2 protein levels comprising the step of administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein modulation of IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to a method of treating a disease or disorder by modulating IKZF2 protein levels comprising the step of administering to a subject in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein modulation of IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to a method of treating a disease or disorder by decreasing IKZF2 protein levels comprising the step of administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein decreasing IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to a method of treating a disease or disorder by decreasing IKZF2 protein levels comprising the step of administering to a subject in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, wherein decreasing IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by modulating IKZF2 protein levels wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by modulating IKZF2 protein levels wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels wherein decreasing IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels wherein decreasing IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by modulating IKZF2 protein levels, wherein modulation of IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by modulating IKZF2 protein levels wherein modulation of IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by decreasing IKZF2 protein levels, wherein decreasing IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by decreasing IKZF2 protein levels wherein decreasing IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a IKZF2-dependent disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a IKZF2-dependent disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a IKZF2-dependent disease or disorder by modulating IKZF2 protein levels, wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a IKZF2-dependent disease or disorder by modulating IKZF2 protein levels, wherein modulation of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels, wherein decreasing IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a IKZF2-dependent disease or disorder by decreasing IKZF2 protein levels, wherein decreasing IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder by modulating IKZF2 protein levels, wherein modulation of IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder by modulating IKZF2 protein levels, wherein modulation of IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder by decreasing IKZF2 protein levels, wherein decreasing IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to the use of a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder by decreasing IKZF2 protein levels, wherein decreasing IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by reducing IKZF2 protein levels wherein reduction of IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by modulating IKZF2 protein levels wherein modulation of IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder by decreasing IKZF2 protein levels wherein decreasing IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats the disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease or disorder by modulating IKZF2 protein levels, wherein modulation of IKZF2 protein levels treats the disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease or disorder by decreasing IKZF2 protein levels, wherein decreasing of IKZF2 protein levels treats the disease or disorder.

In another aspect of the disclosure, the compounds according to the disclosure are formulated into pharmaceutical compositions comprising an effective amount, preferably a pharmaceutically effective amount, of a compound according to the disclosure or salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable excipient or carrier.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating an IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease associated with modulating IKZF2 protein levels.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease associated with the modulation of IKZF2 protein levels.

In some embodiments of the methods disclosed herein, the administration of the compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, is performed orally, parentally, subcutaneously, by injection, or by infusion.

The present disclosure provides degraders of IKZF2 that are therapeutic agents in the treatment of diseases such as cancer and metastasis, in the treatment of diseases affected by the modulation of IKZF2 protein levels, and in the treatment IKZF2-dependent diseases or disorders.

In one embodiment, the disease or disorder that can be treated by the compounds of the present disclosure is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, Ewing's sarcoma non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In one embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST). In another embodiment, the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), and microsatellite stable colorectal cancer (mssCRC). In another embodiment, the IKZF2-dependent disease or disorder is a cancer for which the immune response is deficient or an immunogenic cancer.

The present disclosure provides agents with novel mechanisms of action toward IKZF2 proteins in the treatment of various types of diseases including cancer and metastasis, in the treatment of diseases affected by the modulation of IKZF2 protein levels, and in the treatment IKZF2-dependent diseases or disorders. Ultimately the present disclosure provides the medical community with a novel pharmacological strategy for the treatment of diseases and disorders associated with IKZF2 proteins.

DETAILED DESCRIPTION OF THE DISCLOSURE

The present disclosure relates to compounds and compositions that are capable of modulating IKZF2 protein levels. The disclosure features methods of treating, preventing, or ameliorating a disease or disorder in which IKZF2 plays a role by administering to a patient in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. The methods of the present disclosure can be used in the treatment of a variety of IKZF2-dependent diseases and disorders by modulating IKZF2 protein levels. Modulation of IKZF2 protein levels through degradation provides a novel approach to the treatment, prevention, or amelioration of diseases including, but not limited to, cancer and metathesis, and other IKZF2-dependent diseases or disorders.

In one aspect, the compounds of the disclosure have use as therapeutic agents, particularly for cancers and related diseases. In one aspect, the compounds of the disclosure have IKZF2 degradation activity, preferably having such activity at or below the 50 μM level, and more preferably having such activity at or below the 10 μM level. In another aspect, the compounds of the disclosure have degrader activity for IKZF2 that is selective over one or more of IKZF1, IKZF3, IKZF4, and/or IKZF5. In another aspect, the compounds of the disclosure have degrader activity for both IKZF2 and IKZF4. The compounds of the disclosure have usefulness in treating cancer and other diseases for which such degradation activity would be beneficial for the patient. For example, while not intending to be bound by any theory, the inventors believe that reducing levels of IKZF2 in Tregs in a tumor may allow the patient immune system to more effectively attack the disease. In summary, the present disclosure provides novel IKZF2 degraders useful for the treatment of cancer and other diseases.

In a first aspect of the disclosure, the compounds of Formula (I′) are described:

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof, wherein R_(x), X₁, X₂, and R₁ are as described herein above.

The details of the disclosure are set forth in the accompanying description below. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, illustrative methods and materials are now described. Other features, objects, and advantages of the disclosure will be apparent from the description and from the claims. In the specification and the appended claims, the singular forms also include the plural unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications cited in this specification are incorporated herein by reference in their entireties.

Definition of Terms and Conventions Used

Terms not specifically defined herein should be given the meanings that would be given to them by one of skill in the art in light of the disclosure and the context. As used in the specification and appended claims, however, unless specified to the contrary, the following terms have the meaning indicated and the following conventions are adhered to.

A. Chemical Nomenclature, Terms, and Conventions

In the groups, radicals, or moieties defined below, the number of carbon atoms is often specified preceding the group, for example, (C₁-C₁₀)alkyl means an alkyl group or radical having 1 to 10 carbon atoms. In general, for groups comprising two or more subgroups, the last named group is the radical attachment point, for example, “alkylaryl” means a monovalent radical of the formula alkyl-aryl-, while “arylalkyl” means a monovalent radical of the formula aryl-alkyl-. Furthermore, the use of a term designating a monovalent radical where a divalent radical is appropriate shall be construed to designate the respective divalent radical and vice versa. Unless otherwise specified, conventional definitions of terms control and conventional stable atom valences are presumed and achieved in all formulas and groups. The articles “a” and “an” refer to one or more than one (e.g., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

The term “and/or” means either “and” or “or” unless indicated otherwise.

The term “optionally substituted” means that a given chemical moiety (e.g., an alkyl group) can (but is not required to) be bonded other substituents (e.g., heteroatoms). For instance, an alkyl group that is optionally substituted can be a fully saturated alkyl chain (e.g., a pure hydrocarbon). Alternatively, the same optionally substituted alkyl group can have substituents different from hydrogen. For instance, it can, at any point along the chain be bounded to a halogen atom, a hydroxyl group, or any other substituent described herein. Thus, the term “optionally substituted” means that a given chemical moiety has the potential to contain other functional groups, but does not necessarily have any further functional groups. Suitable substituents used in the optional substitution of the described groups include, without limitation, halogen, oxo, —OH, —CN, —COOH, —CH₂CN, —O—(C₁-C₆)alkyl, (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, —O—(C₂-C₆)alkenyl, —O—(C₂-C₆)alkynyl, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl, —OH, —OP(O)(OH)₂, —OC(O)(C₁-C₆)alkyl, —C(O)(C₁-C₆)alkyl, —OC(O)O(C₁-C₆)alkyl, —NH₂, —NH((C₁-C₆)alkyl), —N((C₁-C₆)alkyl)₂, —NHC(O)(C₁-C₆)alkyl, —C(O)NH(C₁-C₆)alkyl, —S(O)₂(C₁-C₆)alkyl, —S(O)NH(C₁-C₆)alkyl, and S(O)N((C₁-C₆)alkyl)₂. The substituents can themselves be optionally substituted. “Optionally substituted” as used herein also refers to substituted or unsubstituted whose meaning is described below.

The term “substituted” means that the specified group or moiety bears one or more suitable substituents wherein the substituents may connect to the specified group or moiety at one or more positions. For example, an aryl substituted with a cycloalkyl may indicate that the cycloalkyl connects to one atom of the aryl with a bond or by fusing with the aryl and sharing two or more common atoms.

The term “unsubstituted” means that the specified group bears no substituents.

Unless otherwise specifically defined, “aryl” means a cyclic, aromatic hydrocarbon group having 1 to 3 aromatic rings, including monocyclic or bicyclic groups such as phenyl, biphenyl, or naphthyl. When containing two aromatic rings (bicyclic, etc.), the aromatic rings of the aryl group are optionally joined at a single point (e.g., biphenyl), or fused (e.g., naphthyl). The aryl group is optionally substituted by one or more substituents, e.g., 1 to 5 substituents, at any point of attachment. Exemplary substituents include, but are not limited to, —H, -halogen, —CN, —O—(C₁-C₆)alkyl, (C₁-C₆)alkyl, —O—(C₂-C₆)alkenyl, —O—(C₂-C₆)alkynyl, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl, —OH, —OP(O)(OH)₂, —OC(O)(C₁-C₆)alkyl, —C(O)(C₁-C₆)alkyl, —OC(O)O(C₁-C₆) alkyl, —NH₂, —NH((C₁-C₆)alkyl), —N((C₁-C₆)alkyl)₂, —S(O)₂—(C₁-C₆)alkyl, —S(O)NH(C₁-C₆)alkyl, and S(O)N((C₁-C₆)alkyl)₂. The substituents are themselves optionally substituted. Furthermore, when containing two fused rings, the aryl groups optionally have an unsaturated or partially saturated ring fused with a fully saturated ring. Exemplary ring systems of these aryl groups include, but are not limited to, phenyl, biphenyl, naphthyl, anthracenyl, phenalenyl, phenanthrenyl, indanyl, indenyl, tetrahydronaphthalenyl, tetrahydrobenzoannulenyl, and the like.

Unless otherwise specifically defined, “heteroaryl” means a monovalent monocyclic aromatic radical of 5 to 24 ring atoms or a polycyclic aromatic radical, containing one or more ring heteroatoms selected from N, O, or S, the remaining ring atoms being C. Heteroaryl as herein defined also means a bicyclic heteroaromatic group wherein the heteroatom is selected from N, O, or S. The aromatic radical is optionally substituted independently with one or more substituents described herein. Examples include, but are not limited to, furyl, thienyl, pyrrolyl, pyridyl, pyrazolyl, pyrimidinyl, imidazolyl, isoxazolyl, oxazolyl, oxadiazolyl, pyrazinyl, indolyl, thiophen-2-yl, quinolyl, benzopyranyl, isothiazolyl, thiazolyl, thiadiazole, indazole, benzimidazolyl, thieno[3,2-b]thiophene, triazolyl, triazinyl, imidazo[1,2-b]pyrazolyl, furo[2,3-c]pyridinyl, imidazo[1,2-a]pyridinyl, indazolyl, pyrrolo[2,3-c]pyridinyl, pyrrolo[3,2-c]pyridinyl, pyrazolo[3,4-c]pyridinyl, thieno[3,2-c]pyridinyl, thieno[2,3-c]pyridinyl, thieno[2,3-b]pyridinyl, benzothiazolyl, indolyl, indolinyl, indolinonyl, dihydrobenzothiophenyl, dihydrobenzofuranyl, benzofuran, chromanyl, thiochromanyl, tetrahydroquinolinyl, dihydrobenzothiazine, dihydrobenzoxanyl, quinolinyl, isoquinolinyl, 1,6-naphthyridinyl, benzo[de]isoquinolinyl, pyrido[4,3-b][1,6]naphthyridinyl, thieno[2,3-b]pyrazinyl, quinazolinyl, tetrazolo[1,5-a]pyridinyl, [1,2,4]triazolo[4,3-a]pyridinyl, isoindolyl, pyrrolo[2,3-b]pyridinyl, pyrrolo[3,4-b]pyridinyl, pyrrolo[3,2-b]pyridinyl, imidazo[5,4-b]pyridinyl, pyrrolo[1,2-a]pyrimidinyl, tetrahydropyrrolo[1,2-a]pyrimidinyl, 3,4-dihydro-2H-1Δ²-pyrrolo[2,1-b]pyrimidine, dibenzo[b,d]thiophene, pyridin-2-one, furo[3,2-c]pyridinyl, furo[2,3-c]pyridinyl, 1H-pyrido[3,4-b][1,4]thiazinyl, benzooxazolyl, benzoisoxazolyl, furo[2,3-b]pyridinyl, benzothiophenyl, 1,5-naphthyridinyl, furo[3,2-b]pyridine, [1,2,4]triazolo[1,5-a]pyridinyl, benzo[1,2,3]triazolyl, imidazo[1,2-a]pyrimidinyl, [1,2,4]triazolo[4,3-b]pyridazinyl, benzo[c][1,2,5]thiadiazolyl, benzo[c][1,2,5]oxadiazole, 1,3-dihydro-2H-benzo[d]imidazol-2-one, 3,4-dihydro-2H-pyrazolo[1,5-b][1,2]oxazinyl, 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridinyl, thiazolo[5,4 d]thiazolyl, imidazo[2,1-b][1,3,4]thiadiazolyl, thieno[2,3-b]pyrrolyl, 3H-indolyl, and derivatives thereof. Furthermore, when containing two fused rings the aryl groups herein defined may have an unsaturated or partially saturated ring fused with a fully saturated ring. Exemplary ring systems of these heteroaryl groups include indolinyl, indolinonyl, dihydrobenzothiophenyl, dihydrobenzofuran, chromanyl, thiochromanyl, tetrahydroquinolinyl, dihydrobenzothiazine, 3,4-dihydro-1H-isoquinolinyl, 2,3-dihydrobenzofuran, indolinyl, indolyl, and dihydrobenzoxanyl.

Halogen or “halo” mean fluorine, chlorine, bromine, or iodine.

“Alkyl” means a straight or branched chain saturated hydrocarbon containing 1-12 carbon atoms. Examples of a (C₁-C₆)alkyl group include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, neopentyl, and isohexyl.

“Alkoxy” means a straight or branched chain saturated hydrocarbon containing 1-12 carbon atoms containing a terminal “O” in the chain, e.g., —O(alkyl). Examples of alkoxy groups include, without limitation, methoxy, ethoxy, propoxy, butoxy, t-butoxy, or pentoxy groups.

“Alkenyl” means a straight or branched chain unsaturated hydrocarbon containing 2-12 carbon atoms. The “alkenyl” group contains at least one double bond in the chain. The double bond of an alkenyl group can be unconjugated or conjugated to another unsaturated group. Examples of alkenyl groups include ethenyl, propenyl, n-butenyl, isobutenyl, pentenyl, or hexenyl. An alkenyl group can be unsubstituted or substituted and may be straight or branched.

“Alkynyl” means a straight or branched chain unsaturated hydrocarbon containing 2-12 carbon atoms. The “alkynyl” group contains at least one triple bond in the chain. Examples of alkenyl groups include ethynyl, propargyl, n-butynyl, isobutynyl, pentynyl, or hexynyl. An alkynyl group can be unsubstituted or substituted.

“Cycloalkyl” or “carbocyclyl” means a monocyclic or polycyclic saturated carbon ring containing 3-18 carbon atoms. Examples of cycloalkyl groups include, without limitations, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptanyl, cyclooctanyl, norboranyl, norborenyl, bicyclo[2.2.2]octanyl, or bicyclo[2.2.2]octenyl and derivatives thereof. A (C₃-C₅)cycloalkyl is a cycloalkyl group containing between 3 and 8 carbon atoms. A cycloalkyl group can be fused (e.g., decalin) or bridged (e.g., norbomane).

“Heterocyclyl” or “heterocycloalkyl” means a saturated or partially saturated monocyclic or polycyclic ring containing carbon and at least one heteroatom selected from oxygen, nitrogen, or sulfur (O, N, or S) and wherein there is not delocalized n electrons (aromaticity) shared among the ring carbon or heteroatoms. The heterocycloalkyl ring structure may be substituted by one or more substituents. The substituents can themselves be optionally substituted. Examples of heterocyclyl rings include, but are not limited to, oxetanyl, azetadinyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, oxazolinyl, oxazolidinyl, thiazolinyl, thiazolidinyl, pyranyl, thiopyranyl, tetrahydropyranyl, dioxalinyl, piperidinyl, morpholinyl, thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S-dioxide, piperazinyl, azepinyl, oxepinyl, diazepinyl, tropanyl, oxazolidinonyl, 1,4-dioxanyl, dihydrofuranyl, 1,3-dioxolanyl, imidazolidinyl, imidazolinyl, dithiolanyl, and homotropanyl.

“Haloalkyl” means an alkyl group substituted with one or more halogens. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, difluoromethyl, pentafluoroethyl, trichloromethyl, etc.

“Haloalkoxy” means an alkoxy group substituted with one or more halogens. Examples of haloalkyl groups include, but are not limited to, trifluoromethoxy, difluoromethoxy, pentafluoroethoxy, trichloromethoxy, etc.

“Cyano” means a substituent having a carbon atom joined to a nitrogen atom by a triple bond, e.g., C≡N.

“Amino” means a substituent containing at least one nitrogen atom (e.g., —NH₂).

“Hydroxyalkyl” means an alkyl group substituted with one or more —OH groups. Examples of hydroxyalkyl groups include HO—CH₂—, HO—CH₂CH₂—, and CH₂—CH(OH)—.

“Spirocycloalkyl” or “spirocyclyl” means carbogenic bicyclic ring systems with both rings connected through a single atom. The rings can be different in size and nature, or identical in size and nature. Examples include spiropentane, spriohexane, spiroheptane, spirooctane, spirononane, or spirodecane. One or both of the rings in a spirocycle can be fused to another ring carbocyclic, heterocyclic, aromatic, or heteroaromatic ring. A (C₃-C₁₂)spirocycloalkyl is a spirocycle containing between 3 and 12 carbon atoms.

“Spiroheterocycloalkyl” or “spiroheterocyclyl” means a spirocycle wherein at least one of the rings is a heterocycle one or more of the carbon atoms can be substituted with a heteroatom (e.g., one or more of the carbon atoms can be substituted with a heteroatom in at least one of the rings). One or both of the rings in a spiroheterocycle can be fused to another ring carbocyclic, heterocyclic, aromatic, or heteroaromatic ring.

B. Salt, Prodrug, Derivative, and Solvate Terms and Conventions

“Prodrug” or “prodrug derivative” mean a covalently-bonded derivative or carrier of the parent compound or active drug substance which undergoes at least some biotransformation prior to exhibiting its pharmacological effect(s). In general, such prodrugs have metabolically cleavable groups and are rapidly transformed in vivo to yield the parent compound, for example, by hydrolysis in blood, and generally include esters and amide analogs of the parent compounds. The prodrug is formulated with the objectives of improved chemical stability, improved patient acceptance and compliance, improved bioavailability, prolonged duration of action, improved organ selectivity, improved formulation (e.g., increased hydrosolubility), and/or decreased side effects (e.g., toxicity). In general, prodrugs themselves have weak or no biological activity and are stable under ordinary conditions. Prodrugs can be readily prepared from the parent compounds using methods known in the art, such as those described in A Textbook of Drug Design and Development, Krogsgaard-Larsen and H. Bundgaard (eds.), Gordon & Breach, 1991, particularly Chapter 5: “Design and Applications of Prodrugs”; Design of Prodrugs, H. Bundgaard (ed.), Elsevier, 1985; Prodrugs: Topical and Ocular Drug Delivery, K. B. Sloan (ed.), Marcel Dekker, 1998; Methods in Enzymology, K. Widder et al. (eds.), Vol. 42, Academic Press, 1985, particularly pp. 309-396; Burger's Medicinal Chemistry and Drug Discovery, 5th Ed., M. Wolff (ed.), John Wiley & Sons, 1995, particularly Vol. 1 and pp. 172-178 and pp. 949-982; Pro-Drugs as Novel Delivery Systems, T. Higuchi and V. Stella (eds.), Am. Chem. Soc., 1975; Bioreversible Carriers in Drug Design, E. B. Roche (ed.), Elsevier, 1987, each of which is incorporated herein by reference in their entireties.

“Pharmaceutically acceptable prodrug” as used herein means a prodrug of a compound of the disclosure which is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible.

“Salt” means an ionic form of the parent compound or the product of the reaction between the parent compound with a suitable acid or base to make the acid salt or base salt of the parent compound. Salts of the compounds of the present disclosure can be synthesized from the parent compounds, which contain a basic or acidic moiety, by conventional chemical methods. Generally, the salts are prepared by reacting the free base or acid parent compound with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid or base in a suitable solvent or various combinations of solvents.

“Pharmaceutically acceptable salt” means a salt of a compound of the disclosure which is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, generally water or oil-soluble or dispersible, and effective for their intended use. The term includes pharmaceutically-acceptable acid addition salts and pharmaceutically-acceptable base addition salts. As the compounds of the present disclosure are useful in both free base and salt form, in practice, the use of the salt form amounts to use of the base form. Lists of suitable salts are found in, e.g., S. M. Birge et al., J. Pharm. Sci., 1977, 66, pp. 1-19, which is hereby incorporated by reference in its entirety.

“Pharmaceutically-acceptable acid addition salt” means those salts which retain the biological effectiveness and properties of the free bases and which are not biologically or otherwise undesirable, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, nitric acid, phosphoric acid, and the like, and organic acids such as acetic acid, trichloroacetic acid, trifluoroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 2-acetoxybenzoic acid, butyric acid, camphoric acid, camphorsulfonic acid, cinnamic acid, citric acid, digluconic acid, ethanesulfonic acid, glutamic acid, glycolic acid, glycerophosphoric acid, hemisulfic acid, heptanoic acid, hexanoic acid, formic acid, fumaric acid, 2-hydroxyethanesulfonic acid (isethionic acid), lactic acid, maleic acid, hydroxymaleic acid, malic acid, malonic acid, mandelic acid, mesitylenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, nicotinic acid, 2-naphthalenesulfonic acid, oxalic acid, pamoic acid, pectinic acid, phenylacetic acid, 3-phenylpropionic acid, picric acid, pivalic acid, propionic acid, pyruvic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, tartaric acid, p-toluenesulfonic acid, undecanoic acid, and the like.

“Pharmaceutically-acceptable base addition salt” means those salts which retain the biological effectiveness and properties of the free acids and which are not biologically or otherwise undesirable, formed with inorganic bases such as ammonia or hydroxide, carbonate, or bicarbonate of ammonium or a metal cation such as sodium, potassium, lithium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Particularly preferred are the ammonium, potassium, sodium, calcium, and magnesium salts. Salts derived from pharmaceutically-acceptable organic nontoxic bases include salts of primary, secondary, and tertiary amines, quaternary amine compounds, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion-exchange resins, such as methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, TEA, isopropylamine, tripropylamine, tributylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, tetramethylammonium compounds, tetraethylammonium compounds, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, dibenzylamine, N,N-dibenzylphenethylamine, 1-ephenamine, N,N′-dibenzylethylenediamine, polyamine resins, and the like. Particularly preferred organic nontoxic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

“Solvate” means a complex of variable stoichiometry formed by a solute, for example, a compound of Formula (I′) or Formula (I)) and solvent, for example, water, ethanol, or acetic acid. This physical association may involve varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances, the solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. In general, such solvents selected for the purpose of the disclosure do not interfere with the biological activity of the solute. Solvates encompasses both solution-phase and isolatable solvates. Representative solvates include hydrates, ethanolates, methanolates, and the like.

“Hydrate” means a solvate wherein the solvent molecule(s) is/are water.

The compounds of the present disclosure as discussed below include the free base or acid thereof, their salts, solvates, and prodrugs and may include oxidized sulfur atoms or quaternized nitrogen atoms in their structure, although not explicitly stated or shown, particularly the pharmaceutically acceptable forms thereof. Such forms, particularly the pharmaceutically acceptable forms, are intended to be embraced by the appended claims.

C. Isomer Terms and Conventions

“Isomers” means compounds having the same number and kind of atoms, and hence the same molecular weight, but differing with respect to the arrangement or configuration of the atoms in space. The term includes stereoisomers and geometric isomers.

“Stereoisomer” or “optical isomer” mean a stable isomer that has at least one chiral atom or restricted rotation giving rise to perpendicular dissymmetric planes (e.g., certain biphenyls, allenes, and spiro compounds) and can rotate plane-polarized light. Because asymmetric centers and other chemical structure exist in the compounds of the disclosure, which may give rise to stereoisomerism, the disclosure contemplates stereoisomers and mixtures thereof. The compounds of the disclosure and their salts include asymmetric carbon atoms and may therefore exist as single stereoisomers, racemates, and as mixtures of enantiomers and diastereomers. Typically, such compounds will be prepared as a racemic mixture. If desired, however, such compounds can be prepared or isolated as pure stereoisomers, i.e., as individual enantiomers or diastereomers, or as stereoisomer-enriched mixtures. As discussed in more detail below, individual stereoisomers of compounds are prepared by synthesis from optically active starting materials containing the desired chiral centers or by preparation of mixtures of enantiomeric products followed by separation or resolution, such as conversion to a mixture of diastereomers followed by separation or recrystallization, chromatographic techniques, use of chiral resolving agents, or direct separation of the enantiomers on chiral chromatographic columns. Starting compounds of particular stereochemistry are either commercially available or are made by the methods described below and resolved by techniques well-known in the art.

“Enantiomers” means a pair of stereoisomers that are non-superimposable mirror images of each other.

“Diastereoisomers” or “diastereomers” mean optical isomers which are not mirror images of each other.

“Racemic mixture” or “racemate” mean a mixture containing equal parts of individual enantiomers.

“Non-racemic mixture” means a mixture containing unequal parts of individual enantiomers.

“Geometrical isomer” means a stable isomer which results from restricted freedom of rotation about double bonds (e.g., cis-2-butene and trans-2-butene) or in a cyclic structure (e.g., cis-1,3-dichlorocyclobutane and trans-1,3-dichlorocyclobutane). Because carbon-carbon double (olefinic) bonds, C═N double bonds, cyclic structures, and the like may be present in the compounds of the disclosure, the disclosure contemplates each of the various stable geometric isomers and mixtures thereof resulting from the arrangement of substituents around these double bonds and in these cyclic structures. The substituents and the isomers are designated using the cis/trans convention or using the E or Z system, wherein the term “E” means higher order substituents on opposite sides of the double bond, and the term “Z” means higher order substituents on the same side of the double bond. A thorough discussion of E and Z isomerism is provided in J. March, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 4th ed., John Wiley & Sons, 1992, which is hereby incorporated by reference in its entirety. Several of the following examples represent single E isomers, single Z isomers, and mixtures of E/Z isomers. Determination of the E and Z isomers can be done by analytical methods such as x-ray crystallography, ¹H NMR, and ¹³C NMR.

Some of the compounds of the disclosure can exist in more than one tautomeric form. As mentioned above, the compounds of the disclosure include all such tautomers.

It is well-known in the art that the biological and pharmacological activity of a compound is sensitive to the stereochemistry of the compound. Thus, for example, enantiomers often exhibit strikingly different biological activity including differences in pharmacokinetic properties, including metabolism, protein binding, and the like, and pharmacological properties, including the type of activity displayed, the degree of activity, toxicity, and the like. Thus, one skilled in the art will appreciate that one enantiomer may be more active or may exhibit beneficial effects when enriched relative to the other enantiomer or when separated from the other enantiomer. Additionally, one skilled in the art would know how to separate, enrich, or selectively prepare the enantiomers of the compounds of the disclosure from this disclosure and the knowledge of the prior art.

Thus, although the racemic form of drug may be used, it is often less effective than administering an equal amount of enantiomerically pure drug; indeed, in some cases, one enantiomer may be pharmacologically inactive and would merely serve as a simple diluent. For example, although ibuprofen had been previously administered as a racemate, it has been shown that only the S-isomer of ibuprofen is effective as an anti-inflammatory agent (in the case of ibuprofen, however, although the R-isomer is inactive, it is converted in vivo to the S-isomer, thus, the rapidity of action of the racemic form of the drug is less than that of the pure S-isomer). Furthermore, the pharmacological activities of enantiomers may have distinct biological activity. For example, S-penicillamine is a therapeutic agent for chronic arthritis, while R-penicillamine is toxic. Indeed, some purified enantiomers have advantages over the racemates, as it has been reported that purified individual isomers have faster transdermal penetration rates compared to the racemic mixture. See U.S. Pat. Nos. 5,114,946 and 4,818,541.

Thus, if one enantiomer is pharmacologically more active, less toxic, or has a preferred disposition in the body than the other enantiomer, it would be therapeutically more beneficial to administer that enantiomer preferentially. In this way, the patient undergoing treatment would be exposed to a lower total dose of the drug and to a lower dose of an enantiomer that is possibly toxic or an inhibitor of the other enantiomer.

Preparation of pure enantiomers or mixtures of desired enantiomeric excess (ee) or enantiomeric purity are accomplished by one or more of the many methods of (a) separation or resolution of enantiomers, or (b) enantioselective synthesis known to those of skill in the art, or a combination thereof. These resolution methods generally rely on chiral recognition and include, for example, chromatography using chiral stationary phases, enantioselective host-guest complexation, resolution or synthesis using chiral auxiliaries, enantioselective synthesis, enzymatic and nonenzymatic kinetic resolution, or spontaneous enantioselective crystallization. Such methods are disclosed generally in Chiral Separation Techniques: A Practical Approach (2nd Ed.), G. Subramanian (ed.), Wiley-VCH, 2000; T. E. Beesley and R. P. W. Scott, Chiral Chromatography, John Wiley & Sons, 1999; and Satinder Ahuja, Chiral Separations by Chromatography, Am. Chem. Soc., 2000. Furthermore, there are equally well-known methods for the quantitation of enantiomeric excess or purity, for example, GC, HPLC, CE, or NMR, and assignment of absolute configuration and conformation, for example, CD ORD, X-ray crystallography, or NMR.

In general, all tautomeric forms and isomeric forms and mixtures, whether individual geometric isomers or stereoisomers or racemic or non-racemic mixtures, of a chemical structure or compound is intended, unless the specific stereochemistry or isomeric form is specifically indicated in the compound name or structure.

D. Pharmaceutical Administration and Treatment Terms and Conventions

A “patient” or “subject” is a mammal, e.g., a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or nonhuman primate, such as a monkey, chimpanzee, baboon or, rhesus. In certain embodiments, the subject is a primate. In yet other embodiments, the subject is a human.

An “effective amount” or “therapeutically effective amount” when used in connection with a compound means an amount of a compound of the present disclosure that (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein.

The terms “pharmaceutically effective amount” or “therapeutically effective amount” means an amount of a compound according to the disclosure which, when administered to a patient in need thereof, is sufficient to effect treatment for disease-states, conditions, or disorders for which the compounds have utility. Such an amount would be sufficient to elicit the biological or medical response of a tissue, system, or patient that is sought by a researcher or clinician. The amount of a compound of according to the disclosure which constitutes a therapeutically effective amount will vary depending on such factors as the compound and its biological activity, the composition used for administration, the time of administration, the route of administration, the rate of excretion of the compound, the duration of treatment, the type of disease-state or disorder being treated and its severity, drugs used in combination with or coincidentally with the compounds of the disclosure, and the age, body weight, general health, sex, and diet of the patient. Such a therapeutically effective amount can be determined routinely by one of ordinary skill in the art having regard to their own knowledge, the prior art, and this disclosure.

As used herein, the term “pharmaceutical composition” refers to a compound of the disclosure, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, together with at least one pharmaceutically acceptable carrier, in a form suitable for oral or parenteral administration.

“Carrier” encompasses carriers, excipients, and diluents and means a material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting a pharmaceutical agent from one organ, or portion of the body, to another organ, or portion of the body of a subject.

A subject is “in need of” a treatment if such subject would benefit biologically, medically, or in quality of life from such treatment (preferably, a human).

As used herein, the term “inhibit”, “inhibition”, or “inhibiting” refers to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.

As used herein, the term “treat”, “treating”, or “treatment” of any disease or disorder refers to alleviating or ameliorating the disease or disorder (i.e., slowing or arresting the development of the disease or at least one of the clinical symptoms thereof); or alleviating or ameliorating at least one physical parameter or biomarker associated with the disease or disorder, including those which may not be discernible to the patient.

As used herein, the term “prevent”, “preventing”, or “prevention” of any disease or disorder refers to the prophylactic treatment of the disease or disorder; or delaying the onset or progression of the disease or disorder.

“Pharmaceutically acceptable” means that the substance or composition must be compatible chemically and/or toxicologically, with the other ingredients comprising a formulation, and/or the mammal being treated therewith.

“Disorder” means, and is used interchangeably with, the terms disease, condition, or illness, unless otherwise indicated.

“Administer”, “administering”, or “administration” means to either directly administering a disclosed compound or pharmaceutically acceptable salt of the disclosed compound or a composition to a subject, or administering a prodrug derivative or analog of the compound or pharmaceutically acceptable salt of the compound or composition to the subject, which can form an equivalent amount of active compound within the subject's body.

“Prodrug” means a compound which is convertible in vivo by metabolic means (e.g., by hydrolysis) to a disclosed compound.

“Compounds of the present disclosure”, “Compounds of Formula (I′) or Formula (I)”, “compounds of the disclosure”, and equivalent expressions (unless specifically identified otherwise) refer to compounds of Formulae (I), (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig), (Ih), (Ii), (Ij), (Ik), (Il), (Im) (In), (Io), (Ip), (Iq), (Ir), (Is), and (It) as herein described including the tautomers, the prodrugs, salts particularly the pharmaceutically acceptable salts, and the solvates and hydrates thereof, where the context so permits thereof, as well as all stereoisomers (including diastereoisomers and enantiomers), rotamers, tautomers, and isotopically labelled compounds (including deuterium substitutions), as well as inherently formed moieties (e.g., polymorphs, solvates and/or hydrates). For purposes of this disclosure, solvates and hydrates are generally considered compositions. In general and preferably, the compounds of the disclosure and the formulas designating the compounds of the disclosure are understood to only include the stable compounds thereof and exclude unstable compounds, even if an unstable compound might be considered to be literally embraced by the compound formula. Similarly, reference to intermediates, whether or not they themselves are claimed, is meant to embrace their salts and solvates, where the context so permits. For the sake of clarity, particular instances when the context so permits are sometimes indicated in the text, but these instances are purely illustrative and it is not intended to exclude other instances when the context so permits.

“Stable compound” or “stable structure” means a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic or diagnostic agent. For example, a compound that would have a “dangling valency” or is a carbanion is not a compound contemplated by the disclosure.

In a specific embodiment, the term “about” or “approximately” means within 20%, preferably within 10%, and more preferably within 5% of a given value or range.

The yield of each of the reactions described herein is expressed as a percentage of the theoretical yield. “Cancer” means any cancer caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, lymphomas, and the like. For example, cancers include, but are not limited to, mesothelioma, leukemias, and lymphomas such as cutaneous T-cell lymphomas (CTCL), noncutaneous peripheral T-cell lymphomas, lymphomas associated with human T-cell lymphotrophic virus (HTLV) such as adult T-cell leukemia/lymphoma (ATLL), B-cell lymphoma, acute nonlymphocytic leukemias, chronic lymphocytic leukemia, chronic myelogenous leukemia, acute myelogenous leukemia, lymphomas, and multiple myeloma, non-Hodgkin lymphoma, acute lymphatic leukemia (ALL), chronic lymphatic leukemia (CLL), Hodgkin's lymphoma, Burkitt lymphoma, adult T-cell leukemia lymphoma, acute-myeloid leukemia (AML), chronic myeloid leukemia (CML), or hepatocellular carcinoma. Further examples include myelodisplastic syndrome, childhood solid tumors such as brain tumors, neuroblastoma, retinoblastoma, Wilms' tumor, bone tumors, and soft-tissue sarcomas, common solid tumors of adults such as head and neck cancers (e.g., oral, laryngeal, nasopharyngeal, and esophageal), genitourinary cancers (e.g., prostate, bladder, renal, uterine, ovarian, testicular), lung cancer (e.g., small-cell and non-small cell), breast cancer (e.g., triple-negative breast cancer (TNBC)), pancreatic cancer, melanoma, and other skin cancers, stomach cancer, brain tumors, tumors related to Gorlin's syndrome (e.g., medulloblastoma, meningioma, etc.), and liver cancer. Additional exemplary forms of cancer which may be treated by the subject compounds include, but are not limited to, cancer of skeletal or smooth muscle, stomach cancer, cancer of the small intestine, rectum carcinoma, cancer of the salivary gland, endometrial cancer, adrenal cancer, anal cancer, rectal cancer, parathyroid cancer, and pituitary cancer.

Additional cancers that the compounds described herein may be useful in preventing, treating, and studying are, for example, colon carcinoma, familiary adenomatous polyposis carcinoma, and hereditary non-polyposis colorectal cancer, or melanoma. Further, cancers include, but are not limited to, labial carcinoma, larynx carcinoma, hypopharynx carcinoma, tongue carcinoma, salivary gland carcinoma, gastric carcinoma, adenocarcinoma, thyroid cancer (medullary and papillary thyroid carcinoma), renal carcinoma, kidney parenchyma carcinoma, cervix carcinoma, uterine corpus carcinoma, endometrium carcinoma, chorion carcinoma, testis carcinoma, urinary carcinoma, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumors, gall bladder carcinoma, bronchial carcinoma, multiple myeloma, basalioma, teratoma, retinoblastoma, choroidea melanoma, seminoma, rhabdomyosarcoma, craniopharyngeoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, plasmocytoma, non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST).

“Simultaneously” or “simultaneous” when referring to a method of treating or a therapeutic use means with a combination of a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and one or more second agent(s) means administration of the compound and the one or more second agent(s) by the same route and at the same time.

“Separately” or “separate” when referring to a method of treating or a therapeutic use means with a combination of a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and one or more second agent(s) means administration of the compound and the one or more second agent(s) by different routes and at approximately the same time.

By therapeutic administration “over a period of time” means, when referring to a method of treating or a therapeutic use with a combination of a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and one or more second agent(s), administration of the compound and the one or more second agent(s) by the same or different routes and at different times. In some embodiments, the administration of the compound or the one or more second agent(s) occurs before the administration of the other begins. In this way, it is possible to administer a one of the active ingredients (i.e., a compound of the Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or one or more second agent(s)) for several months before administering the other active ingredient or ingredients. In this case, no simultaneous administration occurs. Another therapeutic administration over a period of time consists of the administration over time of the two or more active ingredients of the combination using different frequencies of administration for each of the active ingredients, whereby at certain time points in time simultaneous administration of all of the active ingredients takes place whereas at other time points in time only a part of the active ingredients of the combination may be administered (e.g., for example. a compound of formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and the one or more second agents the therapeutic administration over a period of time could be such that a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, is administered once a day and the one or more second agent(s) is administered once every four weeks.)

The compounds can be administered simultaneously (as a single preparation or separate preparation), sequentially, separately, or over a period of time to the other drug therapy or treatment modality. In general, a combination therapy envisions administration of two or more drugs during a single cycle or course of therapy.

“IKZF2-dependent disease or disorder” means any disease or disorder which is directly or indirectly affected by the modulation of IKZF2 protein levels.

“IKZF4-dependent disease or disorder” means any disease or disorder which is directly or indirectly affected by the modulation of IKZF4 protein levels.

D. Specific Embodiments and Methods for Testing Compounds of Formula (I′) or Formula (I)

The present disclosure relates to compounds or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, or tautomers thereof, capable of modulating IKZF2 protein levels, which are useful for the treatment of diseases and disorders associated with modulation of IKZF2 protein levels. The disclosure further relates to compounds, or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, or tautomers thereof, which are useful for reducing or decreasing IKZF2 protein levels.

In one embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ia):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ib):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In yet another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ic):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Id):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ie):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (If):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ig):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ih):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ii):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ij):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ik):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Il):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Im):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (In):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Io):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ip):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Iq):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Ir):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (Is):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In another embodiment, the compounds of Formula (I′) or Formula (I) have the structure of Formula (It):

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

In some embodiments of the formulae above (e.g., Formulae (I), (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig), (Ih), (Ii), (Ij), (Ik), (Il), (Im), (In), (Io), (p), (Iq), (Ir), (Is), and (It)), R₁ is

In another embodiment, R₁ is

In some embodiments of the formulae above, X₁ is H, (C₁-C₃)alkyl, (C₁-C₆)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₆)haloalkoxy, (C₃-C₇)cycloalkyl, halogen, —CN, —OH, or —NH₂. In another embodiment, X₁ is H, (C₁-C₃)alkyl, (C₁-C₆)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₃)haloalkoxy, halogen, —CN, —OH, or —NH₂. In yet another embodiment, X₁ is H, (C₁-C₄)alkyl, (C₁-C₆)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, (C₃-C₇)cycloalkyl, or halogen. In another embodiment, X₁ is H, (C₁-C₄)alkyl, (C₁-C₆)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, or halogen. In yet another embodiment, X₁ is H, halogen, —CN, —OH, or —NH₂. In another embodiment, X₁ is H, (C₁-C₄)alkyl, (C₁-C₄)haloalkyl, halogen, —CN, —OH, or —NH₂. In yet another embodiment, X₁ is H, (C₁-C₄)alkyl, (C₁-C₄)haloalkyl, halogen, —OH, or —NH₂. In another embodiment, X₁ is H, (C₁-C₄)alkyl, halogen, —OH, or —NH₂. In yet another embodiment, X₁ is H, halogen, —OH, or —NH₂. In another embodiment, X₁ is H.

In some embodiments of the formulae above, X₂ is H, (C₁-C₃)alkyl, (C₁-C₆)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₆)haloalkoxy, (C₃-C₇)cycloalkyl, halogen, —CN, —OH, or —NH₂. In another embodiment, X₂ is H, (C₁-C₃)alkyl, (C₁-C₆)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₃)haloalkoxy, halogen, —CN, —OH, or —NH₂. In yet another embodiment, X₂ is H, (C₁-C₄)alkyl, (C₁-C₆)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, (C₃-C₇)cycloalkyl, or halogen. In another embodiment, X₂ is H, (C₁-C₄)alkyl, (C₁-C₆)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, or halogen. In yet another embodiment, X₂ is H, halogen, —CN, —OH, or —NH₂. In another embodiment, X₂ is H, (C₁-C₄)alkyl, (C₁-C₄)haloalkyl, halogen, —CN, —OH, or —NH₂. In yet another embodiment, X₂ is H, (C₁-C₄)alkyl, (C₁-C₄)haloalkyl, halogen, —OH, or —NH₂. In another embodiment, X₂ is H, (C₁-C₄)alkyl, halogen, —OH, or —NH₂. In yet another embodiment, X₂ is H, halogen, —OH, or —NH₂. In another embodiment, X₂ is H.

In some embodiments of the formulae above, X₁ is H and X₂ is (C₁-C₃)alkyl, (C₁-C₆)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₆)haloalkoxy, (C₃-C₇)cycloalkyl, halogen, —CN, —OH, or —NH₂. In another embodiment, X₁ is (C₁-C₃)alkyl, (C₁-C₆)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₆)haloalkoxy, (C₃-C₇)cycloalkyl, halogen, —CN, —OH, or —NH₂ and X₂ is H. In yet another embodiment, X₁ is H and X₂ is H.

In some embodiments of the formulae above, R_(x) is D. In another embodiment, R_(x) is H.

In some embodiments of the formulae above, each R₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₃)haloalkoxy, halogen, —CN, —OH, or —NH₂. In another embodiment, R₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₃)haloalkoxy, or halogen. In yet another embodiment, R₂ is independently at each occurrence halogen, —CN, —OH, or —NH₂. In another embodiment, R₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)alkoxy, (C₁-C₃)haloalkyl, or (C₁-C₃)haloalkoxy. In yet another embodiment, R₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, halogen, —CN, —OH, or —NH₂. In another embodiment, R₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, halogen, or —CN. In yet another embodiment, R₂ is independently at each occurrence (C₁-C₃)alkyl, halogen, or —CN. In another embodiment, R₂ is independently at each occurrence (C₁-C₃)alkyl or (C₁-C₃)haloalkyl. In yet another embodiment, R₂ is independently at each occurrence (C₁-C₃)alkyl or —CN. In another embodiment, R₂ is independently at each occurrence (C₁-C₃)alkyl. In yet another embodiment, R₂ is independently at each occurrence methyl, ethyl, n-propyl, or isopropyl.

In some embodiments of the formulae above, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 5- or 6-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 5- or 6-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- or 6-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₄-C₇)cycloalkyl. In yet another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₅-C₇)cycloalkyl. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a 5- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together when on adjacent carbon atoms form a phenyl or a 5-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, two R₂ together when on adjacent carbon atoms form a phenyl or a 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together when on adjacent carbon atoms form a phenyl. In yet another embodiment, two R₂ together when on adjacent carbon atoms form a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together when on adjacent carbon atoms form a 5-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, two R₂ together when on adjacent carbon atoms form a 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl, or two R₂ together when on adjacent carbon atoms a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, two R₂ together with the carbon atoms to which they are attached form a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, or two R₂ together when on adjacent carbon atoms a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, or two R₂ together when on adjacent carbon atoms a phenyl. In yet another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, or two R₂ together when on adjacent carbon atoms a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or two R₂ together when on adjacent carbon atoms form a phenyl. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₄-C₇)cycloalkyl or two R₂ together when on adjacent carbon atoms form a phenyl. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a (C₅-C₇)cycloalkyl or two R₂ together when on adjacent carbon atoms form a phenyl.

In another embodiment, two R₂ together with the carbon atoms to which they are attached form a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S or two R₂ together when on adjacent carbon atoms form a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, two R₂ together with the carbon atoms to which they are attached form a 5- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S or two R₂ together when on adjacent carbon atoms form a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₂ together with the carbon atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S or two R₂ together when on adjacent carbon atoms form a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 4- or 5-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen.

In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 4- or 5-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, each R₃ is (C₁-C₆)alkyl, (C₁-C₃)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₃)haloalkoxy, halogen, —OH, or —NH₂. In another embodiment, R₃ is (C₁-C₆)alkyl, (C₁-C₃)alkoxy, (C₁-C₃)haloalkyl, (C₁-C₃)haloalkoxy, or halogen. In another embodiment, R₃ is (C₁-C₆)alkyl, (C₁-C₃)alkoxy, (C₁-C₃)haloalkyl, or (C₁-C₃)haloalkoxy. In another embodiment, R₃ is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, or halogen. In another embodiment, R₃ is halogen, —OH, or —NH₂. In another embodiment, R₃ is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, halogen, —OH, or —NH₂. In another embodiment, R₃ is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, halogen, or —OH. In another embodiment, R₃ is (C₁-C₆)alkyl or (C₁-C₃)haloalkyl. In another embodiment, R₃ is (C₁-C₆)alkyl.

In some embodiments of the formulae above, R₄ is —OR₅. In another embodiment, R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₅ is H, (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₅ is (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, R₅ is H, (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, R₅ is (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, R₅ is H, (C₁-C₃)alkyl, or (C₁-C₃)haloalkyl, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, R₅ is (C₁-C₆)alkyl, or (C₁-C₃)haloalkyl, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₅ is (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, R₅ is H, (C₁-C₃)alkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, R₅ is H, (C₁-C₃)alkyl, (C₃-C₇)cycloalkyl, or 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, R₅ is H, (C₁-C₃)alkyl, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, R₅ is H, (C₁-C₃)alkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, R₅ is H, (C₁-C₃)alkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl. In yet another embodiment, R₅ is H, (C₁-C₃)alkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₅ is H, (C₁-C₃)alkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₆ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In yet another embodiment, R₆ is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂.

In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In yet another embodiment, R₆ is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three R₇.

In another embodiment, R₆ is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In yet another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, or 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl and heterocycloalkyl are optionally substituted with one to four R₁₂. In yet another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the aryl and heteroaryl are optionally substituted with one to four R₁₂.

In another embodiment, R₆ is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three R₇. In yet another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, or 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In yet another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇.

In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₂-C₆)hydroxyalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the heterocycloalkyl and heteroaryl are optionally substituted with one to four R₁₂. In yet another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl and aryl are optionally substituted with one to four R₁₂. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl and aryl are optionally substituted with one to four R₁₂.

In another embodiment, R₆ is H, (C₁-C₆)alkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In yet another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three R₇.

In some embodiments of the formulae above, R_(6′) is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In yet another embodiment, R_(6′) is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂.

In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In yet another embodiment, R_(6′) is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three R₇.

In another embodiment, R_(6′) is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In yet another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂. In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, or 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl and heterocycloalkyl are optionally substituted with one to four R₁₂. In yet another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₂-C₆)hydroxyalkyl, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the aryl and heteroaryl are optionally substituted with one to four R₁₂.

In another embodiment, R_(6′) is (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is substituted with one to three R₇. In yet another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, or 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In yet another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇.

In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₂-C₆)hydroxyalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the heterocycloalkyl and heteroaryl are optionally substituted with one to four R₁₂. In yet another embodiment, R₆ is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl and aryl are optionally substituted with one to four R₁₂. In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl and aryl are optionally substituted with one to four R₁₂. In yet another embodiment, R_(6′) is H or (C₁-C₆)alkyl, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R_(6′) is H or (C₁-C₆)alkyl, wherein the alkyl is substituted with one to three R₇. In yet another embodiment, R_(6′) is H or (C₁-C₆)alkyl.

In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇. In yet another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₁-C₃)haloalkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three R₁₁. In another embodiment, R_(6′) is H, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, or (C₆-C₁₀)aryl, wherein the alkyl is optionally substituted with one to three R₇. In yet another embodiment, R_(6′) is H or (C₁-C₆)alkyl, wherein the alkyl is optionally substituted with one to three R₇. In another embodiment, R_(6′) is H or (C₁-C₆)alkyl, wherein the alkyl is substituted with one to three R₇. In yet another embodiment, R_(6′) is H or (C₁-C₆)alkyl.

In some embodiments of the formulae above, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 8-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. In another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 5- to 8-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 6- to 8-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 7- or 8-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. In another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈. In yet another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 5- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈. In another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- or 5-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈. In yet another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈.

In another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 6- or 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈. In yet another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈. In another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 5-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈. In yet another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈. In another embodiment, R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three R₈.

In some embodiments of the formulae above, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to three substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂. In yet another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In yet another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S.

In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In yet another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S.

In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In yet another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, and halogen. In another embodiment, R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to three R₉. In another embodiment, each R₇ is (C₃-C₇)cycloalkyl or 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl and heterocycloalkyl are optionally substituted with one to three R₉. In yet another embodiment, each R₇ is (C₆-C₁₀)aryl or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the aryl and heteroaryl are optionally substituted with one to three R₉.

In another embodiment, each R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one to three R₉. In yet another embodiment, each R₇ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the heterocycloalkyl and heteroaryl are optionally substituted with one to three R₉. In another embodiment, each R₇ is (C₃-C₇)cycloalkyl or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl and heteroaryl are optionally substituted with one to three R₉. In yet another embodiment, each R₇ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S or (C₆-C₁₀)aryl, wherein the heterocycloalkyl and aryl are optionally substituted with one to three R₉.

In another embodiment, each R₇ is (C₃-C₇)cycloalkyl optionally substituted with one to three R₉. In yet another embodiment, each R₇ is (C₃-C₇)cycloalkyl substituted with one to three R₉. In another embodiment, each R₇ is (C₃-C₇)cycloalkyl. In yet another embodiment, each R₇ is (C₆-C₁₀)aryl optionally substituted with one to three R₉. In another embodiment, each R₇ is (C₆-C₁₀)aryl substituted with one to three R₉. In yet another embodiment, each R₇ is (C₆-C₁₀)aryl.

In another embodiment, each R₇ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, optionally substituted with one to three R₉. In yet another embodiment, each R₇ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, substituted with one to three R₉. In another embodiment, each R₇ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, each R₇ is 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, optionally substituted with one to three R₉. In another embodiment, each R₇ is 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, substituted with one to three R₉. In yet another embodiment, each R₇ is 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, —CN, —OH, —NR₁₃R₁₄, —NH₂, —O(C₃-C₇)cycloalkyl, —O-4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, —O(C₆-C₁₀)aryl, or —O-5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkoxy is optionally substituted with one to three R₁₀ and the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with one to three R₁₁. In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, —CN, —OH, —NR₁₃R₁₄, and —NH₂. In another embodiment, each R₈ is independently at each occurrence —NR₁₃R₁₄, —NH₂, —O(C₃-C₇)cycloalkyl, —O-4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, —O(C₆-C₁₀)aryl, or —O-5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, —CN, —OH, —NR₁₃R₁₄, —O(C₃-C₇)cycloalkyl, —O-4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, —O(C₆-C₁₀)aryl, or —O-5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkoxy is optionally substituted with one or two R₁₀ and the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with one to three R₁.

In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, —CN, —OH, or —NH₂, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, —CN, or —OH, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, halogen, —CN, or —OH, wherein the alkoxy is optionally substituted with (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, and (C₆-C₁₀)aryl.

In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, halogen, —CN, or —OH, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, halogen, —CN, or —OH, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, halogen, —CN, or —OH, wherein the alkoxy is optionally substituted with one to three substituents independently selected from 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, halogen, —CN, or —OH, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl and 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, halogen, —CN, or —OH, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, halogen, —CN, or —OH, wherein the alkoxy is optionally substituted with one to three substituents independently selected from 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₈ is independently at each occurrence halogen, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₄)haloalkoxy, halogen, —CN, or —OH, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl and (C₆-C₁₀)aryl.

In another embodiment, each R₈ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, halogen, —CN, —OH, or —NH₂. In another embodiment, each R₈ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —CN, —OH, or —NH₂. In yet another embodiment, each R₈ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, or halogen. In another embodiment, each R₈ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, or —OH. In yet another embodiment, each R₈ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, or —NH₂. In another embodiment, each R₈ is independently at each occurrence halogen, —CN, —OH, or —NH₂. In yet another embodiment, each R₈ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, or halogen. In another embodiment, each R₈ is independently at each occurrence (C₁-C₆)alkyl or halogen. In another embodiment, each R₈ is independently at each occurrence (C₁-C₆)alkyl, halogen, or —CN. In yet another embodiment, each R₈ is independently at each occurrence halogen or —CN. In yet another embodiment, each R₈ is independently at each occurrence halogen.

In some embodiments of the formulae above, two R₈ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₄-C₆)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₄-C₅)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₄-C₆)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₄-C₅)cycloalkyl or a 6- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₄-C₅)cycloalkyl or a 4- to 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅.

In another embodiment, two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl. In yet another embodiment, two R₈ together with the atoms to which they are attached form a (C₅-C₆)cycloalkyl. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₆-C₇)cycloalkyl. In yet another embodiment, two R₈ together with the atoms to which they are attached form a (C₅)cycloalkyl. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₆)cycloalkyl. In yet another embodiment, two R₈ together with the atoms to which they are attached form a (C₇)cycloalkyl.

In another embodiment, two R₈ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl optionally substituted with two R₁₅. In yet another embodiment, two R₈ together with the atoms to which they are attached form a (C₅-C₆)cycloalkyl optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₆-C₇)cycloalkyl optionally substituted with two R₁₅. In yet another embodiment, two R₈ together with the atoms to which they are attached form a (C₅)cycloalkyl optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a (C₆)cycloalkyl optionally substituted with two R₁₅. In yet another embodiment, two R₈ together with the atoms to which they are attached form a (C₇)cycloalkyl optionally substituted with two R₁₅.

In another embodiment, two R₈ together with the atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In yet another embodiment, two R₈ together with the atoms to which they are attached form a 4- or 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₈ together with the atoms to which they are attached form a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In yet another embodiment, two R₈ together with the atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₈ together with the atoms to which they are attached form a 6- or 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In yet another embodiment, two R₈ together with the atoms to which they are attached form a 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₈ together with the atoms to which they are attached form a 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In yet another embodiment, two R₈ together with the atoms to which they are attached form a 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S.

In another embodiment, two R₈ together with the atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In yet another embodiment, two R₈ together with the atoms to which they are attached form a 4- or 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In yet another embodiment, two R₈ together with the atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a 6- or 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In yet another embodiment, two R₈ together with the atoms to which they are attached form a 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ together with the atoms to which they are attached form a 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In yet another embodiment, two R₈ together with the atoms to which they are attached form a 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅.

In another embodiment, two R₈ when on the same carbon atom together with the atoms to which they are attached form a (C₄-C₇)spirocycloalkyl or a 4- to 7-membered spiroheterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅. In another embodiment, two R₈ when on adjacent atoms together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅.

In another embodiment, two R₈ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₈ when on adjacent atoms together with the atoms to which they are attached form a phenyl or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₈ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl. In another embodiment, two R₈ when on adjacent atoms together with the atoms to which they are attached form a 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₈ when on adjacent atoms together with the atoms to which they are attached form a phenyl.

In another embodiment, two R₈ together with the same atom to which they are attached form a ═(O).

In some embodiments of the formulae above, each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, (C₃-C₆)cycloalkyl, —OH, —CN, —NH₂, or —NR₁₃R₁₄. In another embodiment, R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, (C₃-C₆)cycloalkyl, —OH, —CN, or NR₁₃R₁₄. In another embodiment, each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, or —NH₂. In another embodiment, each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, or —NH₂. In another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, or —CN. In another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, halogen, or —CN. In another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, halogen, or —CN. In yet another embodiment, each R₉ is independently at each occurrence halogen, —OH, —CN, or —NH₂. In another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, or halogen. In another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, or halogen. In another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, or halogen. In yet another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, or —CN. In another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl or (C₁-C₃)haloalkyl. In yet another embodiment, each R₉ is independently at each occurrence halogen or —CN. In another embodiment, each R₉ is independently at each occurrence (C₁-C₃)alkyl or halogen. In yet another embodiment, each R₉ is independently at each occurrence (C₁-C₃)haloalkyl.

In some embodiments of the formulae above, two R₉ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one to three substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one to three substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one to three substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, and —CN. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one to three halogen. In yet another embodiment, two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₅-C₆)cycloalkyl. In yet another embodiment, two R₉ together with the atoms to which they are attached form a (C₆-C₇)cycloalkyl. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₄)cycloalkyl. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₅)cycloalkyl. In yet another embodiment, two R₉ together with the atoms to which they are attached form a (C₆)cycloalkyl. In another embodiment, two R₉ together with the atoms to which they are attached form a (C₇)cycloalkyl.

In another embodiment, two R₉ together with the atoms to which they are attached form a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₉ together with the atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₉ together with the atoms to which they are attached form a 6- or 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₉ together with the atoms to which they are attached form a 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₉ together with the atoms to which they are attached form a 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₉ together with the atoms to which they are attached form a 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S.

In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a phenyl or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl or 5-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a phenyl or 5-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a phenyl or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a phenyl. In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a 5-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, two R₉ when on adjacent atoms together with the atoms to which they are attached form a 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, each R₁₀ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₁₀ is (C₃-C₇)cycloalkyl or 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, each R₁₀ is (C₆-C₁₀)aryl or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S

In another embodiment, each R₁₀ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl. In yet another embodiment, each R₁₀ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₁₀ is (C₃-C₇)cycloalkyl or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, each R₁₀ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S or (C₆-C₁₀)aryl. In yet another embodiment, each R₁₀ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₁₀ is 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, each R₁₀ is (C₃-C₇)cycloalkyl. In yet another embodiment, each R₁₀ is (C₆-C₁₀)aryl.

In some embodiments of the formulae above, each R₁₁ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, or —NH₂. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, or —NH₂. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, or —CN. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, halogen, or —CN. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, halogen, or —CN. In yet another embodiment, each R₁₁ is independently at each occurrence halogen, —OH, —CN, or —NH₂. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, or halogen. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, or halogen. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, or halogen. In yet another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, or —CN. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl or (C₁-C₃)haloalkyl. In yet another embodiment, each R₁₁ is independently at each occurrence halogen or —CN. In another embodiment, each R₁₁ is independently at each occurrence (C₁-C₃)alkyl or halogen.

In some embodiments of the formulae above, each R₁₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, or —NH₂. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, or —NH₂. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, or —CN. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, halogen, or —CN. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, halogen, or —CN. In yet another embodiment, each R₁₂ is independently at each occurrence halogen, —OH, —CN, or —NH₂. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, or halogen. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, (C₁-C₆)alkoxy, or halogen. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, or halogen. In yet another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl, (C₁-C₃)haloalkyl, or —CN. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl or (C₁-C₃)haloalkyl. In yet another embodiment, each R₁₂ is independently at each occurrence halogen or —CN. In another embodiment, each R₁₂ is independently at each occurrence (C₁-C₃)alkyl or halogen.

In some embodiments of the formulae above, two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₆-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₄-C₆)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₄-C₅)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 6- or 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- or 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S.

In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₆-C₇)cycloalkyl. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₄-C₆)cycloalkyl. In another embodiment, two R₁₂ together with the atoms to which they are attached form a (C₄-C₅)cycloalkyl.

In another embodiment, two R₁₂ together with the atoms to which they are attached form a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a 6- or 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₂ together with the atoms to which they are attached form a 4- or 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, R₁₃ is selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₃ is selected from (C₁-C₆)alkyl and (C₁-C₆)haloalkyl. In another embodiment, R₁₃ is selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₃ is selected from (C₃-C₇)cycloalkyl and 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, each R₁₃ is selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S

In another embodiment, R₁₃ is selected from (C₃-C₇)cycloalkyl and (C₆-C₁₀)aryl. In yet another embodiment, R₁₃ is selected from 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₃ is selected from (C₃-C₇)cycloalkyl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, R₁₃ is selected from 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S and (C₆-C₁₀)aryl. In yet another embodiment, R₁₃ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₃ is 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₃ is (C₃-C₇)cycloalkyl. In yet another embodiment, each R₁₃ is (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₁₄ is selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₄ is selected from (C₁-C₆)alkyl and (C₁-C₆)haloalkyl. In another embodiment, R₁₄ is selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₄ is selected from (C₃-C₇)cycloalkyl and 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, R₁₄ is selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S.

In another embodiment, R₁₄ is selected from (C₃-C₇)cycloalkyl and (C₆-C₁₀)aryl. In yet another embodiment, R₁₄ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₄ is selected from (C₃-C₇)cycloalkyl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In yet another embodiment, R₁₄ is selected from 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S and (C₆-C₁₀)aryl. In yet another embodiment, R₁₄ is 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₄ is 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁₄ is (C₃-C₇)cycloalkyl. In yet another embodiment, R₁₄ is (C₆-C₁₀)aryl.

In some embodiments of the formulae above, two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₆-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₄-C₆)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₄-C₅)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 6- or 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- or 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S.

In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₆-C₇)cycloalkyl. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₄-C₆)cycloalkyl. In another embodiment, two R₁₅ together with the atoms to which they are attached form a (C₄-C₅)cycloalkyl.

In another embodiment, two R₁₅ together with the atoms to which they are attached form a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a 6- or 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S. In another embodiment, two R₁₅ together with the atoms to which they are attached form a 4- or 5-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S.

In some embodiments of the formulae above, m is 0, 1, or 2. In another embodiment, m is 0 or 1. In yet another embodiment, m is 1 or 2. In another embodiment, m is 0. In yet another embodiment, m is 1. In another embodiment, m is 2.

In some embodiments of the formulae above, m1 is 0, 1, or 2. In another embodiment, m1 is 0 or 1. In yet another embodiment, m1 is 1 or 2. In another embodiment, m1 is 0. In yet another embodiment, m1 is 1. In another embodiment, m1 is 2.

In some embodiments of the formulae above, n1 is 0, 1, or 2. In another embodiment, n1 is 1, 2, or 3. In another embodiment, n1 is 0 or 1. In another embodiment, n1 is 1 or 2. In another embodiment, n1 is 2 or 3. In another embodiment, n1 is 0. In another embodiment, n1 is 1. In another embodiment, n1 is 2. In another embodiment, n1 is 3.

In some embodiments of the formulae above, each s and n is independently 1, 2, or 3, wherein s+n is ≤4. In another embodiment, each s and n is independently 1 or 2, wherein s+n is ≤4. In another embodiment, each s and n is independently 2 or 3, wherein s+n is ≤4. In another embodiment, s is 1 and n is 1. In another embodiment, s is 2 and n is 2. In another embodiment, s is 1 and n is 2. In another embodiment, s is 2 and n is 1. In another embodiment, s is 3 and n is 1. In another embodiment, s is 1 and n is 3.

In some embodiments of the formulae above, R₁ is

and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

and n1 is 1 or 2.

In some embodiments of the formulae above, R₁ is

n is 1 or 2, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

n1 is 1 or 2, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, n1 is 1 or 2 and R₄ is —OR₅.

In some embodiments of the formulae above, n1 is 1 or 2 and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

n1 is 1 or 2, and m1 is 0.

In some embodiments of the formulae above, R₁ is

n1 is 1 or 2, m1 is 0, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

n1 is 1 or 2, m1 is 0, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, m1 is 0 and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, m1 is 0 and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

and m1 is 0.

In some embodiments of the formulae above, R₁ is

m1 is 0, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

m1 is 0, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, n1 is 1 or 2 and m1 is 0.

In some embodiments of the formulae above, n1 is 1 or 2, m1 is 0, and R₄ is —OR₅.

In some embodiments of the formulae above, n1 is 1 or 2, m1 is 0, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

n1 is 1 or 2, and m1 is 2.

In some embodiments of the formulae above, R₁ is

n1 is 1 or 2, m1 is 2, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

n1 is 1 or 2, m1 is 2, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

and m1 is 2.

In some embodiments of the formulae above, R₁ is

m1 is 2, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

m1 is 2, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, n1 is 1 or 2 and m1 is 2.

In some embodiments of the formulae above, n1 is 1 or 2, m1 is 2, and R₄ is —OR₅.

In some embodiments of the formulae above, n1 is 1 or 2, m1 is 2, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, m1 is 2 and R₄ is —OR₅.

In some embodiments of the formulae above, m1 is 2 and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

n is 2 and s is 1 or 2.

In some embodiments of the formulae above, R₁ is

and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

n is 2, s is 1 or 2, and m is 0 or 1.

In some embodiments of the formulae above, R₁ is

n is 2, s is 1 or 2, m is 0 or 1, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

n is 2, s is 1 or 2, m is 0 or 1, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

n is 2, s is 1 or 2, m is 0 or 1, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

n is 2, s is 1 or 2, m is 0 or 1, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

and m is 0 or 1.

In some embodiments of the formulae above, R₁ is

m is 0 or 1, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

m is 0 or 1, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

m is 0 or 1, and R₄ is —OR₅.

In some embodiments of the formulae above, R₁ is

m is 0 or 1, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, n is 2, s is 1 or 2, and m is 0 or 1.

In some embodiments of the formulae above, n is 2, s is 1 or 2, m is 0 or 1, and R₄ is —OR₅.

In some embodiments of the formulae above, n is 2, s is 1 or 2, m is 0 or 1, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, n is 2, s is 1 or 2 and R₄ is —OR₅.

In some embodiments of the formulae above, n is 2, s is 1 or 2 and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, m is 0 or 1, and R₄ is —OR₅.

In some embodiments of the formulae above, m is 0 or 1, and R₄ is —NR₆R_(6′).

In some embodiments of the formulae above, R₁ is

m1 is 0, R₄ is —OR₅, and R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, and R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl.

In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 0, 1, or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 0. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 0, 1, or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 0. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₈ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 2.

In some embodiments of the formulae above, R₁ is

m1 is 0, R₄ is —OR₅, and R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, and R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl.

In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 0, 1, or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 1 or 2. In another embodiment, R₁ is

R₁ is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 0. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three substituents each independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, and n1 is 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 0, 1, or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 0. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —OR₅, R₈ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl, and n1 is 2.

In some embodiments of the formulae above, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), and R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, and n1 is 0, 1, or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, and n1 is 0. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, and n1 is 2.

In some embodiments of the formulae above, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), and R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. In another embodiment, R is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, and R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 0, 1, or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 0.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, and R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 0, 1, or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 0.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, and R₇ is (C₆-C₁₀)aryl.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl, and n1 is 0, 1, or 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl, and n1 is 0.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈, R₇ is (C₆-C₁₀)aryl, and n1 is 2.

In some embodiments of the formulae above, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), and R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 0, 1, or 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 0.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 0, 1, or 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 0.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H, (C₁-C₆)haloalkyl, or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl, and n1 is 2.

In some embodiments of the formulae above, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉, and n1 is 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl optionally substituted with one or two R₉, and n1 is 2.

In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl, and n1 is 1 or 2. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl, and n1 is 1. In another embodiment, R₁ is

m1 is 0, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R₇ is (C₆-C₁₀)aryl, and n1 is 2.

In some embodiments of the formulae above, R₁ is

m is 0, n is 1, and s is 1. In another embodiment, R₁ is

m is 0, n is 1, s is 1, and R₄ is —OR₅. In another embodiment, R₁ is

m is 0, n is 1, s is 1, R₄ is —OR₅, and R₅ is H, (C₆-C₁₀)aryl, or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl. In another embodiment, R₁ is

m is 0, n is 1, s is 1, R₄ is —OR₅, and R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₁ is

m is 0, n is 1, s is 1, and R₄ is —NR₆R_(6′). In another embodiment, R₁ is

m is 0, n is 1, s is 1, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m is 0, n is 1, s is 1, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m is 0, n is 1, s is 1, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉.

In another embodiment, R₁ is

m is 0, n is 1, s is 1, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl optionally substituted with one or more R₉.

In another embodiment, R₁ is

m is 0, n is 1, s is 1, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₁ is

m is 0, n is 2, and s is 1. In another embodiment, R is

m is 0, n is 2, s is 1, and R₄ is —OR₅. In another embodiment, R₁ is

m is 0, n is 2, s is 1, R₄ is —OR₅, and R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₁ is

m is 0, n is 2, s is 1, and R₄ is —NR₆R_(6′). In another embodiment, R₁ is

m is 0, n is 2, s is 1, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m is 0, n is 2, s is 1, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m is 0, n is 2, s is 1, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉.

In another embodiment, R₁ is

m is 0, n is 2, s is 1, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl optionally substituted with one or more R₉.

In another embodiment, R₁ is

m is 0, n is 2, s is 1, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₁ is

m is 0, n is 2, and s is 2. In another embodiment, R is

n is 2, s is 2, and R₄ is —OR₅. In another embodiment, R₁ is

m is 0, n is 2, s is 2, R₄ is —OR₅, and R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₁ is

m is 0, n is 2, s is 2, and R₄ is —NR₆R_(6′). In another embodiment, R₁ is

m is 0, n is 2, s is 2, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m is 0, n is 2, s is 2, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m is 0, n is 2, s is 2, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉.

In another embodiment, R₁ is

m is 0, n is 2, s is 2, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl optionally substituted with one or more R₉.

In another embodiment, R₁ is

m is 0, n is 2, s is 2, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₁ is

m is 0, n is 2, s is 1 or 2, and R₄ is —OR₅. In another embodiment, R₁ is

m is 0, n is 2, s is 1 or 2, R₄ is —OR₅ and R₅ is H or (C₁-C₆)alkyl optionally substituted with one to three (C₆-C₁₀)aryl.

In some embodiments of the formulae above, R₁ is

m is 0, n is 2, s is 1 or 2, and R₄ is —NR₆R_(6′). In another embodiment, R₁ is

m is 0, n is 2, s is 1 or 2, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m is 0, n is 2, s is 1 or 2, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇. In another embodiment, R₁ is

m is 0, n is 2, s is 1 or 2, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₃-C₇)cycloalkyl or (C₆-C₁₀)aryl, wherein the cycloalkyl and aryl are optionally substituted with one or two R₉.

In another embodiment, R₁ is

m is 0, n is 2, s is 1 or 2, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl optionally substituted with one or more R₉.

In another embodiment, R₁ is

m is 0, n is 2, s is 1 or 2, R₄ is —NR₆R_(6′), R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, and R₇ is (C₆-C₁₀)aryl.

In another embodiment, R₁ is

m1 is 0, n1 is 1, 2, or 3, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. In another embodiment, R is

m1 is 0, n1 is 2 or 3, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₁ is

m1 is 0, n1 is 1 or 2, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₁ is

m1 is 0, n1 is 1, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₁ is

m1 is 0, n1 is 2, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₁ is

m1 is 0, n1 is 3, R₄ is —NR₆R_(6′), and R₆ is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, R_(6′) is H or (C₁-C₆)alkyl optionally substituted with one to three R₇, or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₁ is

m1 is 0, n1 is 1, 2, or 3, R₄ is —NR₆R_(6′), and R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. In yet another embodiment, R₁ is

m1 is 0, n1 is 2 or 3, R₄ is —NR₆R_(6′), and R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₁ is

m1 is 0, n1 is 1 or 2, R₄ is —NR₆R_(6′), and R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. In yet another embodiment, R₁ is

m1 is 0, n1 is 1, R₄ is —NR₆R_(6′), and R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

In another embodiment, R₁ is

m1 is 0, n1 is 2, R₄ is —NR₆R_(6′), and R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈. In yet another embodiment, R₁ is

m1 is 0, n1 is 3, R₄ is —NR₆R_(6′), and R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈.

Embodiment 1

A compound of Formula (I′), wherein:

X₁ and X₂ are each independently H, (C₁-C₄)alkyl, (C₁-C₆)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₆)haloalkoxy, (C₃-C₇)cycloalkyl, halogen, —CN, —OH, or —NH₂;

R_(x) is H or D;

R₁ is

each R₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —CN, —OH, or —NH₂; or

two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₃ is (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —OH, or —NH₂;

R₄ is —OR₅ or —NR₆R_(6′);

R₅ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂; or

R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 8-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₉;

each R₈ is independently at each occurrence halogen, (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, —CN, —OH, —NR₁₃R₁₄, —NH₂, —O(C₃-C₇)cycloalkyl, —O-4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, —O(C₆-C₁₀)aryl, or —O-5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkoxy is optionally substituted with one to three R₁₀ and the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with one to three R₁; or

two R₈ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅; or two R₈ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S; or two R₈ together with the same atom to which they are attached form a ═(O);

each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, or —NH₂; or

two R₉ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one or more substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂; or two R₉ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

each R₁₀ is independently at each occurrence selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

each R₁ is independently at each occurrence selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, and —NH₂;

each R₁₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, or —NH₂;

two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S;

R₁₃ and R₁₄ are each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S;

m and m1 are each independently 0, 1 or 2;

n1 is 0, 1, 2, or 3; and

each s and n is independently 1, 2, or 3, wherein s+n is ≤4;

or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, and tautomer thereof.

Embodiment 2

The compound of Embodiment 1, having a Formula (I), wherein:

R_(x) is H or D;

R₁ is

each R₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —CN, —OH, or —NH₂; or

two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₃ is (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, OH, or —NH₂;

R₄ is —OR₅ or —NR₆R_(6′);

R₅ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇; or

R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₉;

each R₈ is independently at each occurrence halogen, (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, —CN, —OH, or —NH₂, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S; or

two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S;

each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, or —NH₂; or

two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one or more substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

m and m1 are each independently 0, 1, or 2;

n1 is 0, 1, 2, or 3; and

each s and n is independently 1, 2, or 3, wherein s+n is ≤4;

or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, and tautomer thereof.

Embodiment 2A

The compound of Embodiment 1, having a Formula (I), wherein:

R_(x) is H or D;

R₁ is

each R₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —CN, —OH, or —NH₂; or

two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring comprising 1-3 heteroatoms selected from O, N, and S; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₃ is (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —OH, or —NH₂;

R₄ is —OR₅ or —NR₆R_(6′);

R₅ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S;

R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇; or

R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈; or

R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally comprising 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring comprising 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl comprising 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₉;

each R₈ is independently at each occurrence halogen, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —CN, —OH, or —NH₂; or

two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S;

each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, or —NH₂; or

two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring comprising 1-2 heteroatoms selected from O, N, and S optionally substituted with one or more substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂;

m and m1 are each independently 0, 1 or 2;

n1 is 0, 1, 2, or 3; and

each s and n is independently 1, 2, or 3, wherein s+n is ≤4;

or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, and tautomers thereof.

Embodiment 3

The compound of any one of Embodiments 1, 2, and 2A, wherein R₁ is

Embodiment 4

The compound of any one of Embodiments 1, 2, and 2A, wherein n1 is 0, 1, or 2.

Embodiment 5

The compound of any one of Embodiments 1 to 3 and 2A, wherein m is 0.

Embodiment 6

The compound of any one of Embodiments 1 to 3 and 2A, wherein m is 2.

Embodiment 7

The compound of any one of Embodiments 1, 2, and 2A, wherein R₁ is

Embodiment 8

The compound of Embodiment 7, wherein n is 2 and s is 1 or 2.

Embodiment 9

The compound of Embodiment 7 or 8, wherein m is 0 or 1.

Embodiment 10

The compound of any one of Embodiments 1, 2, and 2A, having a Formula (Ia) or Formula (Ib), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, and tautomer thereof.

Embodiment 11

The compound of any one of Embodiments 1, 2, and 2A, having a Formula (Ic), Formula (Id), Formula (Ie), Formula (If), Formula (Ig), Formula (Ih), Formula (Ii), or Formula (Ij), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, and tautomer thereof.

Embodiment 12

The compound of any one of Embodiments 1, 2, and 2A, having a Formula (Ik), Formula (Il), Formula (Im), Formula (In), Formula (Io), or Formula (Ip), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, and tautomer thereof.

Embodiment 13

The compound of any one of Embodiments 1 to 12 and 2A, wherein R₄ is —OR₅.

Embodiment 14

The compound of any one of Embodiments 1 to 12 and 2A, wherein R₄ is —NR₆R_(6′).

Embodiment 15

The compound of any one of Embodiments 1, 2, and 2A, having a Formula (Iq), Formula (Ir), Formula (Is), or Formula (It), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, and tautomer thereof.

Embodiment 16

The compound of Embodiment 1 selected from:

Compound Structure/Compound Name

or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Embodiment 17

A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

Embodiment 18

The pharmaceutical composition of Embodiment 17 further comprising at least one additional pharmaceutical agent.

Embodiment 19

The pharmaceutical composition of Embodiment 17 or Embodiment 18 for use in the treatment of a disease or disorder that is affected by the reduction of IKZF2 protein levels.

Embodiment 20

A method of degrading IKZF2 comprising administering to the patient in need thereof a compound of any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Embodiment 21

A method of treating a disease or disorder that is affected by the modulation of IKZF2 protein levels comprising administering to the patient in need thereof a compound of any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Embodiment 22

A method of modulating IKZF2 protein levels comprising administering to the patient in need thereof a compound of any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Embodiment 23

A method of reducing the proliferation of a cell the method comprising, contacting the cell with a compound of any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and reducing IKZF2 protein levels.

Embodiment 24

A method of treating cancer comprising administering to the patient in need thereof a compound of any one of Embodiments 1-15 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Embodiment 25

The method of Embodiment 24, wherein the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma.

Embodiment 26

The method of Embodiment 24, wherein the cancer is a cancer for which the immune response is deficient or an immunogenic cancer.

Embodiment 27

A method for reducing IKZF2 protein levels in a subject comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt.

Embodiment 28

The method of Embodiments any one of 20-27, wherein administering is performed orally, parentally, subcutaneously, by injection, or by infusion.

Embodiment 29

A compound according to any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a disease or disorder that is affected by the reduction of IKZF2 protein levels.

Embodiment 30

Use of a compound according to any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder that is affected by the reduction of IKZF2 protein levels.

Embodiment 31

A compound according to any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease or disorder associated with the reduction of IKZF2 protein levels.

Embodiment 32

The compound of Embodiment 31, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma.

Embodiment 33

Use of a compound according to any one of Embodiments 1-16 and 2A, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease or disorder associated with the reduction of IKZF2 protein levels.

Embodiment 34

The use of Embodiment 33, wherein the disease or disorder is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, and Ewing's sarcoma.

Embodiment 35

The method of Embodiment 24, wherein the cancer is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST).

Embodiment 36

The compound of Embodiment 31, wherein the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST).

Embodiment 37

The use of Embodiment 33, wherein the disease or disorder is selected from non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST).

In another embodiment of the disclosure, the compounds of the present disclosure are enantiomers. In some embodiments the compounds are the (S)-enantiomer. In other embodiments, the compounds are the (R)-enantiomer. In yet other embodiments, the compounds of the present disclosure may be (+) or (−) enantiomers.

It should be understood that all isomeric forms are included within the present disclosure, including mixtures thereof. If the compound contains a double bond, the substituent may be in the E or Z configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis- or trans-configuration. All tautomeric forms are also intended to be included.

Compounds of the disclosure, and pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, and prodrugs thereof may exist in their tautomeric form (for example, as an amide or imino ether). All such tautomeric forms are contemplated herein as part of the present disclosure.

The compounds of the disclosure may contain asymmetric or chiral centers and, therefore, exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the disclosure as well as mixtures thereof, including racemic mixtures, form part of the present disclosure. In addition, the present disclosure embraces all geometric and positional isomers. For example, if a compound of the disclosure incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the disclosure. Each compound herein disclosed includes all the enantiomers that conform to the general structure of the compound. The compounds may be in a racemic or enantiomerically pure form, or any other form in terms of stereochemistry. The assay results may reflect the data collected for the racemic form, the enantiomerically pure form, or any other form in terms of stereochemistry.

Diastereomeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods well known to those skilled in the art, such as, for example, by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Also, some of the compounds of the disclosure may be atropisomers (e.g., substituted biaryls) and are considered as part of this disclosure. Enantiomers can also be separated by use of a chiral HPLC column.

It is also possible that the compounds of the disclosure may exist in different tautomeric forms, and all such forms are embraced within the scope of the disclosure and chemical structures and names. Also, for example, all keto-enol and imine-enamine forms of the compounds are included in the disclosure.

All stereoisomers (for example, geometric isomers, optical isomers, and the like) of the present compounds (including those of the salts, solvates, esters, and prodrugs of the compounds as well as the salts, solvates and esters of the prodrugs), such as those which may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms, are contemplated within the scope of this disclosure, as are positional isomers (such as, for example, 4-pyridyl and 3-pyridyl). (For example, if a compound of Formula (I′) or Formula (I) incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the disclosure. Also, for example, all keto-enol and imine-enamine forms of the compounds are included in the disclosure.) Individual stereoisomers of the compounds of the disclosure may, for example, be substantially free of other isomers, or is admixed, for example, as racemates or with all other, or other selected, stereoisomers.

The chiral centers of the compounds of the disclosure can have the S or R configuration as defined by the IUPAC 1974 Recommendations. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R)- or (S)-configuration. Substituents at atoms with unsaturated double bonds may, if possible, be present in cis-(Z)- or trans-(E)-form.

The use of the terms “salt”, “solvate”, “ester,” “prodrug”, and the like, is intended to equally apply to the salt, solvate, ester, and prodrug of enantiomers, stereoisomers, rotamers, tautomers, positional isomers, racemates, or prodrugs of the inventive compounds.

The compounds of the disclosure may form salts which are also within the scope of this disclosure. Reference to a compound of the Formula herein is generally understood to include reference to salts thereof, unless otherwise indicated.

The compounds and intermediates may be isolated and used as the compound per se. Any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into compounds of the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and, such as ²H, ³H, ¹¹C, ¹³C, ⁴C, ¹⁵N, ¹⁸F, ³¹P, ³²P, respectively. The disclosure includes various isotopically labeled compounds as defined herein, for example those into which radioactive isotopes, such as ³H, ¹³C, and ¹⁴C, are present. Such isotopically labelled compounds are useful in metabolic studies (with ¹⁴C), reaction kinetic studies (with, for example ²H or ³H), detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays, or in radioactive treatment of patients. In particular, an ¹⁸F, ¹¹C or labeled compound may be particularly desirable for PET or SPECT studies.

Further, substitution with heavier isotopes, particularly deuterium (i.e., ²H or D) may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life, reduced dosage requirements, reduced CYP450 inhibition (competitive or time dependent) or an improvement in therapeutic index. For example, substitution with deuterium may modulate undesirable side effects of the undeuterated compound, such as competitive CYP450 inhibition, time dependent CYP450 inactivation, etc. It is understood that deuterium in this context is regarded as a substituent in compounds of the present disclosure. The concentration of such a heavier isotope, specifically deuterium, may be defined by the isotopic enrichment factor. The term “isotopic enrichment factor” as used herein means the ratio between the isotopic abundance and the natural abundance of a specified isotope. If a substituent in a compound of this disclosure is denoted deuterium, such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

Isotopically-labeled compounds of the present disclosure can generally be prepared by conventional techniques known to those skilled in the art or by carrying out the procedures disclosed in the schemes or in the examples and preparations described below using an appropriate isotopically-labeled reagent in place of the non-isotopically labeled reagent.

Pharmaceutically acceptable solvates in accordance with the disclosure include those wherein the solvent of crystallization may be isotopically substituted, e.g., D₂O, d₆-acetone, d₆-DMSO.

The present disclosure relates to compounds which are modulators of IKZF2 protein levels. In one embodiment, the compounds of the present disclosure decrease IKZF2 protein levels. In yet one embodiment, the compounds of the present disclosure reduce IKZF2 protein levels. In another embodiment, the compounds of the present disclosure are degraders of IKZF2.

The present disclosure relates to compounds which are modulators of IKZF2 and IKZF4 protein levels. In one embodiment, the compounds of the present disclosure decrease IKZF2 and IKZF4 protein levels. In yet one embodiment, the compounds of the present disclosure reduce IKZF2 and IKZF4 protein levels. In another embodiment, the compounds of the present disclosure are degraders of IKZF2.

In some embodiments, the compounds of the disclosure are selective over other proteins. As used herein “selective modulator”, “selective degrader”, or “selective compound” means, for example, a compound of the disclosure, that effectively modulates, decreases, or reduces the levels of a specific protein or degrades a specific protein to a greater extent than any other protein. A “selective modulator”, “selective degrader”, or “selective compound” can be identified, for example, by comparing the ability of a compound to modulate, decrease, or reduce the levels of or to degrade a specific protein to its ability to modulate, decrease, or reduce the levels of or to degrade other proteins. In some embodiments, the selectivity can be identified by measuring the EC₅₀ or IC₅₀ of the compounds.

In some embodiments, the compounds of the present application are selective IKZF2 modulators. As used herein “selective IKZF2 modulator”, “selective IKZF2 degrader”, or “selective IKZF2 compound” refers to a compound of the application, for example, that effectively modulates, decrease, or reduces the levels of IKZF2 protein or degrades IKZF2 protein to a greater extent than any other protein, particularly any protein (transcription factor) from the Ikaros protein family (e.g., IKZF1, IKZF3, IKZF4, and IKZF5).

A “selective IKZF2 modulator”, “selective IKZF2 degrader”, or “selective IKZF2 compound” can be identified, for example, by comparing the ability of a compound to modulate IKZF2 protein levels to its ability to modulate levels of other members of the Ikaros protein family or other proteins. For example, a substance may be assayed for its ability to modulate IKZF2 protein levels, as well as IKZF1, IKZF3, IKZF4, IKZF5, and other proteins. In some embodiments, the selectivity can be identified by measuring the EC₅₀ of the compounds. In some embodiments, a selective IKZF2 degrader is identified by comparing the ability of a compound to degrade IKZF2 to its ability to degrade other members of the Ikaros protein family or other proteins.

In certain embodiments, the compounds of the application are IKZF2 degraders that exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 over other proteins (e.g., IKZF1, IKZF3, IKZF4, and IKZF5). In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 over other proteins.

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 over the other members of the Ikaros protein family (e.g., IKZF1, IKZF3, IKZF4, and IKZF5). In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 over the other members of the Ikaros protein family (e.g., IKZF1, IKZF3, IKZF4, and IKZF5).

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 over IKZF1. In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 over IKZF1.

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 over IKZF3. In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 over IKZF3.

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 over IKZF4. In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 over IKZF4.

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 over IKZF5. In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 over IKZF5.

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 and IKZF4 over the other members of the Ikaros protein family (e.g., IKZF1, IKZF3, and IKZF5). In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 and IKZF4 over the other members of the Ikaros protein family (e.g., IKZF1, IKZF3, and IKZF5).

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 and IKZF4 over IKZF1. In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 and IKZF4 over IKZF1.

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 and IKZF4 over IKZF3. In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 and IKZF4 over IKZF3.

In certain embodiments, the compounds of the application exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, or 100-fold selectivity for the degradation of IKZF2 and IKZF4 over IKZF5. In various embodiments, the compounds of the application exhibit up to 1000-fold selectivity for the degradation of IKZF2 and IKZF4 over IKZF5.

In some embodiments, the degradation of IKZF2 is measured by EC₅₀.

Potency of can be determined by EC₅₀ value. A compound with a lower EC₅₀ value, as determined under substantially similar degradation conditions, is a more potent degrader relative to a compound with a higher EC₅₀ value. In some embodiments, the substantially similar conditions comprise determining degradation of protein levels in cells expressing the specific protein, or a fragment of any thereof.

The disclosure is directed to compounds as described herein and pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, or tautomers thereof, and pharmaceutical compositions comprising one or more compounds as described herein, or pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers, or tautomers thereof.

E. Methods of Synthesizing Compounds of Formula (I′) or Formula (I)

The compounds of the present disclosure may be made by a variety of methods, including standard chemistry. Suitable synthetic routes are depicted in the Schemes given below.

The compounds of the present disclosure may be prepared by methods known in the art of organic synthesis as set forth in part by the following synthetic schemes. In the schemes described below, it is well understood that protecting groups for sensitive or reactive groups are employed where necessary in accordance with general principles or chemistry. Protecting groups are manipulated according to standard methods of organic synthesis (T. W. Greene and P. G. M. Wuts, “Protective Groups in Organic Synthesis”, Third edition, Wiley, New York 1999). These groups are removed at a convenient stage of the compound synthesis using methods that are readily apparent to those skilled in the art. The selection processes, as well as the reaction conditions and order of their execution, shall be consistent with the preparation of compounds of Formula (I′) or Formula (I).

Those skilled in the art will recognize if a stereocenter exists in the compounds of the present disclosure. Accordingly, the present disclosure includes all possible stereoisomers (unless specified in the synthesis) and includes not only racemic compounds but the individual enantiomers and/or diastereomers as well. When a compound is desired as a single enantiomer or diastereomer, it may be obtained by stereospecific synthesis or by resolution of the final product or any convenient intermediate. Resolution of the final product, an intermediate, or a starting material may be affected by any suitable method known in the art. See, for example, “Stereochemistry of Organic Compounds” by E. L. Eliel, S. H. Wilen, and L. N. Mander (Wiley-Interscience, 1994).

The compounds described herein may be made from commercially available starting materials or synthesized using known organic, inorganic, and/or enzymatic processes.

Preparation of Compounds

The compounds of the present disclosure can be prepared in a number of ways well known to those skilled in the art of organic synthesis. By way of example, compounds of the present disclosure can be synthesized using the methods described below, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereon as appreciated by those skilled in the art. Preferred methods include but are not limited to those methods described below.

Compounds of the present disclosure can be synthesized by following the steps outlined in General Schemes I, II, III, IV, and V which comprise different sequences of assembling intermediates I-a, I-b, I-c, I-d, I-e, I-f, II-a, II-b, II-c, III-a, III-b, IV-a, IV-b, IV-c, IV-d1, IV-d2, IV-e1, IV-e2, IV-f, V-a, V-b, V-c1, V-c2, V-d1, and V-d2. Starting materials are either commercially available or made by known procedures in the reported literature or as illustrated.

The general way of preparing compounds of Formula (I′) wherein R₁ is

R₄ is —NR₆R_(6′), and R₆ is alkyl substituted with R₇ or optionally substituted alkyl using intermediates I-a, I-b, I-c, I-d, I-e, and I-f is outlined in General Scheme I. Coupling of I-a with 1,2-aminoalcohol I-b using a catalyst (e.g., Ni(glyme)Cl₂ with 4,4-di-tert-butyl-2,2′-dipyridyl (dtbbpy)) and photocatalyst (e.g., 4,4′-bis(1,1-dimethylethyl)-2,2′-bipyridine-N1,N1′]bis[3,5-difluoro-2-[5-(trifluoromethyl)-2-pyridinyl-N]phenyl-C]Iridium(III)hexafluorophosphate (Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆)), a base (e.g., 2,2,6,6-tetramethylpiperidine (TMP), quinuclidine or potassium carbonate (K₂CO₃)), and in a solvent, (e.g., acetonitrile (MeCN)), under irradiation of blue LED light yields I-c. When P is a protecting group (e.g. tert-butyloxycarbonyl (Boc) or 2-(trimethylsilyl)ethoxymethyl (SEM)), intermediate I-c is deprotected using a strong acid such as trifluoroacetic acid (TFA), methanesulfonic acid (MsOH), or hydrochloric acid (HCl) in a solvent (e.g., tetrahydrofuran (THF), MeCN, or dichloromethane (DCM)) optionally at elevated temperature to provide I-d. Reductive amination of I-d with aldehyde or ketone I-f provides compounds of Formula (I′) where R₆ is a substituted alkyl. Alternatively, compounds of Formula (I′) wherein R₆ is an optionally substituted alkyl can be obtained by alkylation of I-d with alkyl halide, tosylate or mesylate I-e in the presence of a base (e.g., triethylamine (Et₃N), N,N-diisopropylethylamine (i-Pr₂NEt), cesium carbonate (Cs₂CO₃), etc.), in a solvent (e.g., MeCN, N,N-dimethylformamide (DMF), etc.), and optionally at elevated temperature.

The general way of preparing compounds of Formula (I′) wherein R₁ is

R₄ is —NR₆R_(6′), and R₆ is alkyl substituted with R₇ or optionally substituted alkyl using intermediates I-a, II-a, II-b, II-c, I-e, and I-f is outlined in General Scheme II. Coupling of I-a with 1,2-aminoalcohol II-a using a catalyst (e.g., Ni(glyme)Cl₂ with 4,4-di-tert-butyl-2,2′-dipyridyl (dtbbpy)) and a photocatalyst (e.g., 4,4′-bis(1,1-dimethylethyl)-2,2′-bipyridine-N1,N1′]bis[3,5-difluoro-2-[5-(trifluoromethyl)-2-pyridinyl-N]phenyl-C]Iridium(III) hexafluorophosphate(Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆)), a base (e.g., TMP, quinuclidine or K₂CO₃), and in a solvent, (e.g., MeCN), under irradiation of blue LED light yields II-b. When P is a protecting group (e.g., Boc or SEM), intermediate II-b is deprotected using a strong acid such as TFA, MsOH, or HCl in a solvent (e.g., THF, MeCN, or DCM) optionally at elevated temperature to provide II-c. Reductive amination of II-c with aldehyde or ketone I-f provides compounds of Formula (I′) where R₆ is a substituted alkyl. Alternatively, compounds of Formula (I′) wherein R₆ is an optionally substituted alkyl can be obtained by alkylation of II-c with alkyl halide, tosylate or mesylate I-e in the presence of a base (e.g., Et₃N, i-Pr₂NEt, Cs₂CO₃, etc.), in a solvent (e.g., MeCN, DMF, etc.), and optionally at elevated temperature.

The general way of preparing compounds of Formula (I′) wherein R₁ is

and R₄ is —OR₅, using intermediates I-a, III-a, and III-b is outlined in General Scheme III. Coupling of I-a with 1,2-diols or 1,2-etheralcohols III-a using a catalyst (e.g., Ni(glyme)Cl₂ with 4,4-di-tert-butyl-2,2′-dipyridyl (dtbbpy)) and a photocatalyst (e.g., 4,4′-bis(1,1-dimethylethyl)-2,2′-bipyridine-N1,N1′]bis[3,5-difluoro-2-[5-(trifluoromethyl)-2-pyridinyl-N]phenyl-C]Iridium(III)hexafluorophosphate (Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆)), a base (e.g., TMP, quinuclidine or K₂CO₃), and in a solvent, (e.g., MeCN), under irradiation of blue LED light yields III-b. When P is a protecting group (e.g., SEM), intermediate III-b is deprotected using a strong acid such as TFA, MsOH, or HCl in a solvent (e.g., THF, MeCN, or DCM) optionally at elevated temperature to provide the desired compounds of Formula (I′) wherein R₅ is H or substituted alkyl.

The general way of preparing compounds of Formula (I′) wherein R₁ is

R₄ is —NR₆R_(6′), and R₆ is alkyl substituted with R₇ or optionally substituted alkyl using intermediates IV-a, IV-b, IV-c, IV-d1, IV-d2, IV-e1, IV-e2, IV-f, I-e, and I-f is outlined in General Scheme IV. Coupling of IV-a with 1,2-aminoalcohol II-a using a catalyst (e.g., Ni(glyme)Cl₂ with 4,4-di-tert-butyl-2,2′-dipyridyl (dtbbpy)) and a photocatalyst, (e.g., 4,4′-bis(1,1-dimethylethyl)-2,2′-bipyridine-N1,N1′]bis[3,5-difluoro-2-[5-(trifluoromethyl)-2-pyridinyl-N]phenyl-C]Iridium(III) hexafluorophosphate(Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆)), a base (e.g., TMP, quinuclidine, or K₂CO₃), and in a solvent, (e.g., MeCN), under irradiation of blue LED light yields IV-b. When P is a protecting group (e.g., Boc), intermediate IV-b is deprotected using a strong acid such as TFA or HCl in a solvent (e.g. THF or DCM) optionally at elevated temperature yields IV-c. Reductive amination of IV-c with aldehyde or ketone I-f provides IV-d. Alternatively, compounds IV-d can be obtained by alkylation of IV-c with an alkyl halide, tosylate or mesylate I-e in the presence of a base (e.g., Et₃N, i-Pr₂NEt, Cs₂CO₃, etc.), in a solvent (e.g., MeCN, DMF, etc.), and optionally at elevated temperature. Intermediate IV-d can then be converted to the corresponding haloester intermediate IV-e using SOCl₂ in a solvent (e.g., EtOH) at elevated temperatures. Cyclization of IV-e with 3-aminopiperidine-2,6-dione IV-f (or its HCl or CF₃CO₂H salt) with base (e.g., i-Pr₂NEt) in a solvent (e.g., DMF) at elevated temperature provides compounds of Formula (I′) wherein R₆ is a substituted alkyl.

The general way of preparing compounds of Formula (I′) wherein R₁ is

R₄ is —NR₆R_(6′), and R₆ is alkyl substituted with R₇ or optionally substituted alkyl using intermediates IV-a, IV-f, V-a, V-b, V-c1, V-c2, V-d1, V-d2, I-b, I-e, and I-f is outlined in General Scheme V. Coupling of IV-a with 1,2-aminoalcohol I-b using a catalyst (e.g., Ni(glyme)Cl₂ with 4,4-di-tert-butyl-2,2′-dipyridyl (dtbbpy)) and a photocatalyst, (e.g., 4,4′-bis(1,1-dimethylethyl)-2,2′-bipyridine-N1,N1′]bis[3,5-difluoro-2-[5-(trifluoromethyl)-2-pyridinyl-N]phenyl-C]Iridium(III)hexafluorophosphate (Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆)), a base (e.g., TMP, quinuclidine, or K₂CO₃), and in a solvent, (e.g., MeCN), under irradiation of blue LED light yields V-a. When P is a protecting group (e.g., Boc), intermediate V-a is deprotected using a strong acid such as TFA or HCl in a solvent (e.g., THF or DCM) optionally at elevated temperature yields V-b. Reductive amination of V-b with aldehyde or ketone I-f provides V-c2. V-c1 can be obtained by alkylation of V-b with an alkyl halide, tosylate or mesylate I-e in the presence of a base (e.g., Et₃N, i-Pr₂NEt, Cs₂CO₃, etc.), in a solvent (e.g., MeCN, DMF, etc.), and optionally at elevated temperature. Intermediates V-c1 and V-c2 can then be converted to the corresponding haloester intermediate V-d1 and Vd2 using SOCl₂ in a solvent (e.g., EtOH) and optionally at elevated temperatures. Cyclization of V-d1 or Vd2 with 3-aminopiperidine-2,6-dione IV-f (or its HCl or CF₃CO₂H salt) using a base (e.g., i-Pr₂NEt) in a solvent (e.g., DMF) and optionally at elevated temperature provides compounds of Formula (I′) wherein R₆ is a substituted alkyl or an optionally substituted alkyl.

A mixture of enantiomers, diastereomers, and cis/trans isomers resulting from the process described above can be separated into their single components by chiral salt technique, chromatography using normal phase, reverse phase or chiral column, depending on the nature of the separation.

Any resulting racemates of compounds of the present disclosure or of intermediates can be resolved into the optical antipodes by known methods, e.g., by separation of the diastereomeric salts thereof, obtained with an optically active acid or base, and liberating the optically active acidic or basic compound. In particular, a basic moiety may thus be employed to resolve the compounds of the present disclosure into their optical antipodes, e.g., by fractional crystallization of a salt formed with an optically active acid, e.g., tartaric acid, dibenzoyl tartaric acid, diacetyl tartaric acid, di-O,O′-p-toluoyl tartaric acid, mandelic acid, malic acid, or camphor-10-sulfonic acid. Racemic compounds of the present disclosure or racemic intermediates can also be resolved by chiral chromatography, e.g., high pressure liquid chromatography (HPLC) using a chiral adsorbent.

Any resulting mixtures of stereoisomers can be separated on the basis of the physicochemical differences of the constituents, into the pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and/or fractional crystallization.

It should be understood that in the description and formula shown above, the various groups R_(x), R₂, R₃, R₅, R_(6′), R₇, X₁, X₂, m, m1, n, n1, and s and other variables are as defined above, except where otherwise indicated. Furthermore, for synthetic purposes, the compounds of General Schemes I, II, III, IV, and V are merely representative with elected radicals to illustrate the general synthetic methodology of the compounds of Formula (I′) as defined herein.

F. Methods of Using Compounds of Formula (I′) or Formula (I)

Another aspect of the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder in a patient associated with modulation of IKZF2 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders associated with modulation of IKZF2 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by the modulation of IKZF2 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders affected by the modulation of IKZF2 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by the reduction of IKZF2 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders affected by the reduction of IKZF2 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by a decrease in IKZF2 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders affected by the reduction or decrease of IKZF2 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for the treatment, prevention, inhibition or elimination of a disease or disorder that is affected by the modulation of IKZF2 protein levels.

In another aspect, the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for the treatment, prevention, inhibition or elimination of a disease or disorder that is affected by the reduction of or a decrease in IKZF2 protein levels.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by the modulation of IKZF2 protein levels.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by the reduction of or a decrease in IKZF2 protein levels.

In another aspect, the present disclosure is directed to a method of modulating IKZF2 protein levels. The method involves administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF2 protein levels are modulated through degradation of the IKZF2 protein. In other embodiments, IKZF2 protein levels are modulated through degradation of the IKZF2 protein mediated by an E3 ligase.

Another aspect of the present disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder in a patient associated with the reduction of or decrease in IKZF2 protein levels, the method comprising administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

The present disclosure also relates to the use of a degrader of IKZF2 for the preparation of a medicament used in the treatment, prevention, inhibition or elimination of a IKZF2-dependent disease or disorder, wherein the medicament comprises a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a method for treating, preventing, inhibiting, or eliminating a IKZF2-dependent disease or disorder, wherein the medicament comprises a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a method for the manufacture of a medicament for treating, preventing, inhibiting, or eliminating a IKZF2-dependent disease or disorder mediated, wherein the medicament comprises a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease or disorder associated with the modulation of IKZF2 protein levels. In some embodiments, IKZF2 levels are modulated through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are modulated through degradation of the IKZF2 protein mediated by an E3 ligase.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating a disease associated with the modulation of IKZF2 protein levels. In some embodiments, IKZF2 levels are modulated through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are modulated through degradation of the IKZF2 protein mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease associated with the modulation of IKZF2 protein levels. In some embodiments, IKZF2 protein levels are modulated through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are modulated through degradation of the IKZF2 protein mediated by an E3 ligase.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease or disorder associated with the reduction of IKZF2 protein levels. In some embodiments, IKZF2 levels are reduced through degradation of the IKZF2 protein. In some embodiments, IKZF2 levels are reduced through degradation of the IKZF2 protein mediated by an E3 ligase.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating a disease associated with the reduction of IKZF2 protein levels. In some embodiments, IKZF2 levels are reduced through degradation of the IKZF2 protein. In some embodiments, IKZF2 levels are reduced through degradation of the IKZF2 protein mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease associated with the reduction of IKZF2 protein levels. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein. In some embodiments, IKZF2 levels are reduced through degradation of the IKZF2 protein mediated by an E3 ligase.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease or disorder associated with a decrease in IKZF2 protein levels. In some embodiments, IKZF2 levels are decreased through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are decreased through degradation of the IKZF2 protein mediated by an E3 ligase.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating a disease associated with a decrease in IKZF2 protein levels. In some embodiments, IKZF2 levels are decreased through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are decreased through degradation of the IKZF2 protein mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease associated with a decrease in IKZF2 protein levels. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are decreased through degradation of the IKZF2 protein mediated by an E3 ligase.

In another aspect, the present disclosure relates to a method of inhibiting IKZF2 activity through degradation of IKZF2. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for inhibiting IKZF2 activity through degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the inhibition of IKZF2 activity through degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for inhibiting IKZF2 activity through degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a method of inhibiting IKZF2 and IKZF4 activity through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for inhibiting IKZF2 and IKZF4 activity through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the inhibition of IKZF2 and IKZF4 activity through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for inhibiting IKZF2 and IKZF4 activity through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for inhibiting IKZF2 and IKZF4 activity through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder associated with modulation of IKZF2 and IKZF4 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders associated with modulation of IKZF2 and IKZF4 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure is directed to a method of modulating IKZF2 and IKZF4 protein levels. The method involves administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF2 and IKZF4 protein levels are modulated through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are modulated through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

Another aspect of the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder associated with modulation of, reduction of, or a decrease in IKZF4 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders associated with modulation of, reduction of, or decrease in IKZF4 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 proteins. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 protein mediated by an E3 ligase.

In another aspect, the present disclosure is directed to a method of modulating, reducing or decreasing IKZF4 protein levels. The method involves administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 proteins. In other embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 protein mediated by an E3 ligase.

Another aspect of the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating, preventing, inhibiting, or eliminating a disease or disorder associated with modulation of, reduction of, or a decrease in IKZF4 protein levels. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 proteins. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 protein mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating, preventing, inhibiting, or eliminating a disease or disorder associated with modulation of, reduction of, or a decrease in IKZF4 protein levels. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 proteins. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 protein mediated by an E3 ligase.

In another aspect, the present disclosure is directed to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating, preventing, inhibiting, or eliminating a disease or disorder associated with modulation of, reduction of, or a decrease in IKZF4 protein levels. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 proteins. In some embodiments, IKZF4 protein levels are modulated, reduced, or decreased through degradation of the IKZF4 protein mediated by an E3 ligase.

Another aspect of the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder associated with the reduction of IKZF2 and IKZF4 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders associated with reduction of IKZF2 and IKZF4 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure is directed to a method of reducing IKZF2 and IKZF4 protein levels. The method involves administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

Another aspect of the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder associated with a decrease in IKZF2 and IKZF4 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders associated with a decrease of IKZF2 and IKZF4 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure is directed to a method of decreasing IKZF2 and IKZF4 protein levels. The method involves administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF2 and IKZF4 protein levels are decreased through degradation of the IKZF2 and IKZF4 proteins. In some embodiments, IKZF2 and IKZF4 protein levels are decreased through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

Another aspect of the present disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder in a patient associated with the modulation of IKZF2 and IKZF4 protein levels, the method comprising administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In one embodiment, the disease or disorder is selected from the group consisting of cancer and metastasis.

In another aspect, the present disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder in a patient associated with the reduction of IKZF2 and IKZF4 protein levels, the method comprising administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In one embodiment, the disease or disorder is selected from the group consisting of cancer and metastasis.

Another aspect of the present disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder in a patient associated with a decrease in IKZF2 and IKZF4 protein levels, the method comprising administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In one embodiment, the disease or disorder is selected from the group consisting of cancer and metastasis.

The present disclosure also relates to the use of a modulator of IKZF2 and IKZF4 protein levels for the preparation of a medicament used in the treatment, prevention, inhibition or elimination of a IKZF2 and IKZF4-dependent disease or disorder, wherein the medicament comprises a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In another aspect, the present disclosure relates to a method for the manufacture of a medicament for treating, preventing, inhibiting, or eliminating a IKZF2 and IKZF4-dependent disease or disorder, wherein the medicament comprises a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease associated with the modulation of IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are modulated through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are modulated through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating a disease associated with the modulation of IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are modulated through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are modulated through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease associated with the reduction of IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating a disease associated with the reduction of IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 are reduced through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a disease associated with a decrease in IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are decreased through degradation of the IKZF2 and IKZF4 proteins. In some embodiments, IKZF2 and IKZF4 protein levels are decreased through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating a disease associated with a decrease in IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 are decreased through degradation of the IKZF2 and IKZF4 proteins. In some embodiments, IKZF2 and IKZF4 protein levels are decreased through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of a disease associated with the modulation of IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are modulated through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are modulated through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder by reducing or decreasing IKZF2 protein levels, wherein reduction or decrease of IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of an IKZF2-dependent disease or disorder by reducing or decreasing IKZF2 protein levels wherein reduction of or decrease in IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

In another aspect, the present disclosure the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating an IKZF2-dependent disease or disorder by reducing or decreasing IKZF2 protein levels wherein reduction of or decrease in IKZF2 protein levels treats the IKZF2-dependent disease or disorder.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2 and IKZF4-dependent disease or disorder by reducing or decreasing IKZF2 and IKZF4 protein levels wherein the reduction of or decrease in IKZF2 and IKZF4 protein levels treats the IKZF2 and IKZF4-dependent disease or disorder.

In another aspect, the present disclosure the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of an IKZF2 and IKZF4-dependent disease or disorder by reducing or decreasing IKZF2 and IKZF4 protein levels wherein the reduction of or decrease in IKZF2 and IKZF4 protein levels treats the IKZF2 and IKZF4-dependent disease or disorder.

In another aspect, the present disclosure the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating an IKZF2 and IKZF4-dependent disease or disorder by reducing or decreasing IKZF2 and IKZF4 protein levels wherein the reduction of or decrease in IKZF2 and IKZF4 protein levels treats the IKZF2 and IKZF4-dependent disease or disorder.

Another aspect of the disclosure relates to a method of treating cancer. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of treating cancer.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating cancer.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of cancer.

Another aspect of the disclosure relates to a method of treating an IKZF2-dependent cancer. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of treating an IKZF2-dependent cancer.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating an IKZF2-dependent cancer.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent cancer.

Another aspect of the disclosure relates to a method of treating an IKZF2-dependent and IKZF4-dependent cancer. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of treating an IKZF2-dependent and IKZF4-dependent cancer.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating an IKZF2-dependent and IKZF4-dependent cancer.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent and IKZF4-dependent cancer.

Another aspect of the disclosure relates to a method of treating a cancer affected by the modulation of, the reduction of, or a decrease in IKZF2 protein levels. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of treating a cancer affected by the modulation of, the reduction of, or a decrease in IKZF2 protein levels

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a cancer affected by the modulation of, the reduction of, or a decrease in IKZF2 protein levels.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a cancer affected by the modulation of, the reduction of, or a decrease in IKZF2 protein levels.

Another aspect of the disclosure relates to a method of treating a cancer affected by the modulation of, the reduction of, or a decrease in IKZF2 and IKZF4 protein levels. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the treatment of treating a cancer affected by the modulation of, the reduction of, or a decrease in IKZF2 and IKZF4 protein levels.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating a cancer affected by the modulation of, the reduction of, or a decrease in IKZF2 and IKZF4 protein levels.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of a cancer affected by the modulation of, the reduction of, or a decrease in IKZF2 and IKZF4 protein levels.

Another aspect of the disclosure relates to a method of degrading IKZF2. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for degrading IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the degradation IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for degrading IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a method of modulating IKZF2 protein levels through degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for modulating IKZF2 protein levels through degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the modulation IKZF2 protein levels through degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for modulating IKZF2 protein levels through degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a method of treating an IKZF2-dependent disease or disorder in a patient in need thereof by modulating IKZF2 protein levels through the degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating an IKZF2-dependent disease or disorder in a patient in need thereof by modulating IKZF2 protein levels through the degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating an IKZF2-dependent disease or disorder in a patient in need thereof, by modulating IKZF2 protein levels through the degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating an IKZF2-dependent disease or disorder in a patient in need thereof by modulating IKZF2 protein levels through the degradation of IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a method of degrading IKZF2. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for degrading IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in degrading IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for degrading IKZF2. In some embodiments, IKZF2 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a method of reducing the proliferation of a cell, the method comprising contacting the cell with a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, that reduces IKZF2 protein levels. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for reducing the proliferation of a cell by reducing IKZF2 protein levels. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in reducing the proliferation of a cell by IKZF2 protein levels. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for reducing the proliferation of a cell by reducing IKZF2 protein levels. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein. In some embodiments, IKZF2 protein levels are reduced through degradation of the IKZF2 protein mediated by an E3 ligase.

In another aspect, the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by the modulation of IKZF2 and IKZF4 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders affected by the modulation of IKZF2 and IKZF4 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the disclosure relates to a method of treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by the reduction of or a decrease in IKZF2 and IKZF4 protein levels. The method comprises administering to a patient in need of a treatment for diseases or disorders affected by the reduction or decrease of IKZF2 and IKZF4 protein levels an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for the treatment, prevention, inhibition or elimination of a disease or disorder that is affected by the modulation of IKZF2 and IKZF4 protein levels.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by the modulation of IKZF2 and IKZF4 protein levels.

In another aspect, the disclosure relates to the use a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for the treatment, prevention, inhibition or elimination of a disease or disorder that is affected by the reduction of or a decrease in IKZF2 and IKZF4 protein levels.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating, preventing, inhibiting, or eliminating a disease or disorder that is affected by the reduction of or a decrease in IKZF2 and IKZF4 protein levels.

Another aspect of the disclosure relates to a method of degrading IKZF2 and IKZF4. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for degrading IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the degradation IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for degrading IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a method of modulating IKZF2 and IKZF4 protein levels through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for modulating IKZF2 and IKZF4 protein levels through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the modulation of IKZF2 and IKZF4 protein levels through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for modulating IKZF2 and IKZF4 protein levels through degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a method of treating an IKZF2-dependent and IKZF4-dependent disease or disorder in a patient in need thereof by modulating IKZF2 and IKZF4 protein levels through the degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating an IKZF2-dependent and IKZF4-dependent disease or disorder in a patient in need thereof by modulating IKZF2 and IKZF4 protein levels through the degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in treating an IKZF2-dependent and IKZF4-dependent disease or disorder in a patient in need thereof by modulating IKZF2 and IKZF4 protein levels through the degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating an IKZF2-dependent or IKZF4-dependent disease or disorder in a patient in need thereof by modulating IKZF2 and IKZF4 protein levels through the degradation of IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a method of degrading IKZF2 and IKZF4. The method comprises administering to a patient in need thereof an effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for degrading IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in degrading IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for degrading IKZF2 and IKZF4. In some embodiments, IKZF2 and IKZF4 protein degradation is mediated by an E3 ligase.

Another aspect of the disclosure relates to a method of reducing the proliferation of a cell, the method comprising contacting the cell with a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and reducing IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

In another aspect, the present disclosure relates to the use a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for reducing the proliferation of a cell by reducing IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in reducing the proliferation of a cell by reducing IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for reducing the proliferation of a cell by reducing IKZF2 and IKZF4 protein levels. In some embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins. In other embodiments, IKZF2 and IKZF4 protein levels are reduced through degradation of the IKZF2 and IKZF4 proteins mediated by an E3 ligase.

In another aspect, the present disclosure relates to a method for treating an IKZF2-dependent disease or disorder. The method comprises the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating an IKZF2-dependent disease or disorder.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating an IKZF2-dependent disease or disorder.

In another aspect, the present disclosure relates to a method for treating an IKZF2-dependent and IKZF4-dependent disease or disorder. The method comprises the step of administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the treatment of an IKZF2-dependent and IKZF4-dependent disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating an IKZF2-dependent and IKZF4-dependent disease or disorder.

Another aspect of the disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for use in the manufacture of a medicament for treating an IKZF2-dependent and IKZF4-dependent disease or disorder.

In another aspect, the present disclosure relates to a method of reducing IKZF2 protein levels. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a method of reducing IKZF2 and IKZF4 protein levels. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof for use in the reduction of IKZF2 protein levels.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof for use in the reduction of IKZF2 and IKZF4 protein levels.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition, in the manufacture of a medicament for reducing IKZF2 protein levels.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for reducing IKZF2 and IKZF4 protein levels.

In another aspect, the present disclosure relates to a method of reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats or ameliorates the disease or disorder. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a method of reducing IKZF2 and IKZF4 protein levels, wherein reduction of IKZF2 and IKZF4 protein levels treats or ameliorates the disease or disorder. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof for use in the reduction of IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats or ameliorates the disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof for use in the reduction of IKZF2 and IKZF4 protein levels, wherein reduction of IKZF2 and IKZF4 protein levels treats or ameliorates the disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition, in the manufacture of a medicament for reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats or ameliorates the disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for reducing IKZF2 and IKZF4 protein levels, wherein reduction of IKZF2 and IKZF4 protein levels treats or ameliorates the disease or disorder.

In another aspect, the present disclosure relates to a method of treating a disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats or ameliorates the disease or disorder. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

Another aspect of the present disclosure relates to a method of treating a disease or disorder by reducing IKZF2 and IKZF4 protein levels, wherein reduction of IKZF2 and IKZF4 protein levels treats or ameliorates the disease or disorder. The method comprises administering to the patient in need thereof a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

In another aspect, the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof for use in the treatment of a disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats or ameliorates the disease or disorder.

Another aspect of the present disclosure relates to a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof for use in the treatment of a disease or disorder by reducing IKZF2 and IKZF4 protein levels, wherein reduction of IKZF2 and IKZF4 protein levels treats or ameliorates the disease or disorder.

In another aspect, the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition, in the manufacture of a medicament for treating a disease or disorder by reducing IKZF2 protein levels, wherein reduction of IKZF2 protein levels treats or ameliorates the disease or disorder.

Another aspect of the present disclosure relates to the use of a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof or a composition comprising a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, in the manufacture of a medicament for treating a disease or disorder by reducing IKZF2 and IKZF4 protein levels, wherein reduction of IKZF2 and IKZF4 protein levels treats or ameliorates the disease or disorder.

The compounds of the present disclosure present disclosure can be used for the treatment, of cancers including, but not limited to, liposarcoma, neuroblastoma, glioblastoma, bladder cancer, adrenocortical cancer, multiple myeloma, colorectal cancer, non-small cell lung cancer, Human Papilloma Virus-associated cervical, oropharyngeal, penis, anal, thyroid, or vaginal cancer or Epstein-Barr Virus-associated nasopharyngeal carcinoma, gastric cancer, rectal cancer, thyroid cancer, Hodgkin lymphoma or diffuse large B-cell lymphoma. In one embodiment, the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, cancer for which the immune response is deficient, an immunogenic cancer, Ewing's sarcoma, non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST).

In some embodiments of the methods above, the IKZF2-dependent disease or disorder is a disease or disorder including, but not limited to, liposarcoma, neuroblastoma, glioblastoma, bladder cancer, adrenocortical cancer, multiple myeloma, colorectal cancer, non-small cell lung cancer, Human Papilloma Virus-associated cervical, oropharyngeal, penis, anal, thyroid, or vaginal cancer or Epstein-Barr Virus-associated nasopharyngeal carcinoma, gastric cancer, rectal cancer, thyroid cancer, Hodgkin lymphoma or diffuse large B-cell lymphoma. In one embodiment, the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, cancer for which the immune response is deficient, an immunogenic cancer, Ewing's sarcoma, non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST).

In some embodiments of the methods above, the disease or disorder affected by the modulation, reduction or decrease of IKZF2 and/or IKZF4 protein levels is a disease or disorder including, but not limited to, liposarcoma, neuroblastoma, glioblastoma, bladder cancer, adrenocortical cancer, multiple myeloma, colorectal cancer, non-small cell lung cancer, Human Papilloma Virus-associated cervical, oropharyngeal, penis, anal, thyroid, or vaginal cancer or Epstein-Barr Virus-associated nasopharyngeal carcinoma, gastric cancer, rectal cancer, thyroid cancer, Hodgkin lymphoma or diffuse large B-cell lymphoma. In one embodiment, the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, cancer for which the immune response is deficient, an immunogenic cancer, Ewing's sarcoma, non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST).

In some embodiments of the methods above, the IKZF2-dependent cancer and IKZF2-dependent and IKZF4-dependent cancer is a cancer selected from liposarcoma, neuroblastoma, glioblastoma, bladder cancer, adrenocortical cancer, multiple myeloma, colorectal cancer, non-small cell lung cancer, Human Papilloma Virus-associated cervical, oropharyngeal, penis, anal, thyroid, or vaginal cancer or Epstein-Barr Virus-associated nasopharyngeal carcinoma, gastric cancer, rectal cancer, thyroid cancer, Hodgkin lymphoma or diffuse large B-cell lymphoma. In one embodiment, the cancer is selected from prostate cancer, breast carcinoma, lymphomas, leukaemia, myeloma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiform, glioma, thyroid cancer, parathyroid tumor, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, and soft tissue sarcomas selected from rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, rhabdoid cancers, cancer for which the immune response is deficient, an immunogenic cancer, Ewing's sarcoma, non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, acute myelogenous leukemia, and gastrointestinal stromal tumor (GIST).

In some embodiments of the methods above, IKZF2 protein levels are modulated by degradation of IKZF2. In some embodiments of the methods above, IKZF2 protein levels are reduced by degradation of IKZF2. In some embodiments of the methods above, IKZF2 protein levels are decreased by degradation of IKZF2.

In some embodiments of the methods above, IKZF2 and IKZF4 protein levels are modulated by degradation of IKZF2 and IKZF4. In some embodiments of the methods above, IKZF2 and IKZF4 protein levels are reduced by degradation of IKZF2 and IKZF4. In some embodiments of the methods above, IKZF2 protein levels are decreased by degradation of IKZF2 and IKZF4.

One therapeutic use of the compounds or compositions of the present disclosure, which modulate IKZF2 and/or IKZF4 protein levels by degradation of IKZF2 and/or IKZF4, is to provide treatment to patients or subjects suffering from cancer and metastasis.

The disclosed compounds of the disclosure can be administered in effective amounts to treat or prevent a disorder and/or prevent the development thereof in subjects.

Compounds of the application can be administered in therapeutically effective amounts in a combinational therapy with one or more therapeutic agents (pharmaceutical combinations) or modalities, e.g., non-drug therapies. For example, synergistic effects can occur with other anti-proliferative, anti-cancer, immunomodulatory or anti-inflammatory substances. Where the compounds of the application are administered in conjunction with other therapies, dosages of the co-administered compounds will of course vary depending on the type of co-drug employed, on the specific drug employed, on the condition being treated and so forth.

Combination therapy includes the administration of the subject compounds in further combination with other biologically active ingredients (such as, but not limited to, a second and different antineoplastic agent or a second agent that targets Helios or another cancer target) and non-drug therapies (such as, but not limited to, surgery or radiation treatment). For instance, the compounds of the application can be used in combination with other pharmaceutically active compounds, preferably compounds that are able to enhance the effect of the compounds of the application. The compounds of the application can be administered simultaneously (as a single preparation or separate preparation) or sequentially to the other drug therapy or treatment modality. In general, a combination therapy envisions administration of two or more drugs during a single cycle or course of therapy.

G. Administration, Pharmaceutical Compositions, and Dosing of Compounds of Formula (I′) or Formula (I)

Administration of the disclosed compounds can be accomplished via any mode of administration for therapeutic agents. These modes include systemic or local administration such as oral, nasal, parenteral, transdermal, subcutaneous, vaginal, buccal, rectal or topical administration modes.

Depending on the intended mode of administration, the disclosed compositions can be in solid, semi-solid or liquid dosage form, such as, for example, injectables, tablets, suppositories, pills, time-release capsules, elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, or the like, sometimes in unit dosages and consistent with conventional pharmaceutical practices. Likewise, they can also be administered in intravenous (both bolus and infusion), intraperitoneal, subcutaneous or intramuscular form, and all using forms well known to those skilled in the pharmaceutical arts.

Illustrative pharmaceutical compositions are tablets and gelatin capsules comprising a compound of the disclosure and a pharmaceutically acceptable carrier, such as a) a diluent, e.g., purified water, triglyceride oils, such as hydrogenated or partially hydrogenated vegetable oil, or mixtures thereof, corn oil, olive oil, sunflower oil, safflower oil, fish oils, such as EPA or DHA, or their esters or triglycerides or mixtures thereof, omega-3 fatty acids or derivatives thereof, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, sodium, saccharin, glucose and/or glycine; b) a lubricant, e.g., silica, talcum, stearic acid, its magnesium or calcium salt, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and/or polyethylene glycol; for tablets also; c) a binder, e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, magnesium carbonate, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, waxes, and/or polyvinylpyrrolidone, if desired; d) a disintegrant, e.g., starches, agar, methyl cellulose, bentonite, xanthan gum, algic acid or its sodium salt, or effervescent mixtures; e) absorbent, colorant, flavorant and sweetener; f) an emulsifier or dispersing agent, such as Tween 80, Labrasol, HPMC, DOSS, caproyl 909, labrafac, labrafil, peceol, transcutol, capmul MCM, capmul PG-12, captex 355, gelucire, vitamin E TGPS or other acceptable emulsifier; and/or g) an agent that enhances absorption of the compound such as cyclodextrin, hydroxypropyl-cyclodextrin, PEG400, PEG200.

Liquid, particularly injectable, compositions can, for example, be prepared by dissolution, dispersion, etc. For example, the disclosed compound is dissolved in or mixed with a pharmaceutically acceptable solvent such as, for example, water, saline, aqueous dextrose, glycerol, ethanol, and the like, to thereby form an injectable isotonic solution or suspension. Proteins such as albumin, chylomicron particles, or serum proteins can be used to solubilize the disclosed compounds.

The disclosed compounds can be also formulated as a suppository that can be prepared from fatty emulsions or suspensions; using polyalkylene glycols such as propylene glycol, as the carrier.

The disclosed compounds can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, containing cholesterol, stearylamine or phosphatidylcholines.

In some embodiments, a film of lipid components is hydrated with an aqueous solution of drug to a form lipid layer encapsulating the drug, as described in U.S. Pat. No. 5,262,564, which is hereby incorporated by reference in its entirety.

Disclosed compounds can also be delivered by the use of monoclonal antibodies as individual carriers to which the disclosed compounds are coupled. The disclosed compounds can also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspanamidephenol, or polyethyleneoxidepolylysine substituted with palmitoyl residues. Furthermore, the disclosed compounds can be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and cross-linked or amphipathic block copolymers of hydrogels. In one embodiment, disclosed compounds are not covalently bound to a polymer, e.g., a polycarboxylic acid polymer, or a polyacrylate.

Parental injectable administration is generally used for subcutaneous, intramuscular or intravenous injections and infusions. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions or solid forms suitable for dissolving in liquid prior to injection.

Another aspect of the disclosure is directed to pharmaceutical compositions comprising a compound of Formula (I′) or Formula (I) and a pharmaceutically acceptable carrier. The pharmaceutical acceptable carrier may further include an excipient, diluent, or surfactant.

Compositions can be prepared according to conventional mixing, granulating or coating methods, respectively, and the present pharmaceutical compositions can contain from about 0.1% to about 99%, from about 5% to about 90%, or from about 1% to about 20% of the disclosed compound by weight or volume.

In one embodiment, the disclosure provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of the present disclosure. In one embodiment, the kit comprises means for separately retaining said compositions, such as a container, divided bottle, or divided foil packet. An example of such a kit is a blister pack, as typically used for the packaging of tablets, capsules and the like.

The kit of the disclosure may be used for administering different dosage forms, for example, oral and parenteral, for administering the separate compositions at different dosage intervals, or for titrating the separate compositions against one another. To assist compliance, the kit of the disclosure typically comprises directions for administration.

The dosage regimen utilizing the disclosed compound is selected in accordance with a variety of factors including type, species, age, weight, sex, and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal or hepatic function of the patient; and the particular disclosed compound employed. A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition.

Effective dosage amounts of the disclosed compounds, when used for the indicated effects, range from about 0.5 mg to about 5000 mg of the disclosed compound as needed to treat the condition. Compositions for in vivo or in vitro use can contain about 0.5, 5, 20, 50, 75, 100, 150, 250, 500, 750, 1000, 1250, 2500, 3500, or 5000 mg of the disclosed compound, or, in a range of from one amount to another amount in the list of doses. In one embodiment, the compositions are in the form of a tablet that can be scored.

H. Combination Therapy

The compounds of the disclosure can be administered in therapeutically effective amounts in a combinational therapy with one or more therapeutic agents (pharmaceutical combinations) or modalities, e.g., non-drug therapies. For example, synergistic effects can occur with other cancer agents. Where the compounds of the application are administered in conjunction with other therapies, dosages of the co-administered compounds will of course vary depending on the type of co-drug employed, on the specific drug employed, on the condition being treated and so forth.

The compounds can be administered simultaneously (as a single preparation or separate preparation), sequentially, separately, or over a period of time to the other drug therapy or treatment modality. In general, a combination therapy envisions administration of two or more drugs during a single cycle or course of therapy. A therapeutic agent is, for example, a chemical compound, peptide, antibody, antibody fragment or nucleic acid, which is therapeutically active or enhances the therapeutic activity when administered to a patient in combination with a compound of the present disclosure.

In one aspect, a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure can be combined with other therapeutic agents, such as other anti-cancer agents, anti-allergic agents, anti-nausea agents (or anti-emetics), pain relievers, cytoprotective agents, and combinations thereof.

In some embodiments, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof of the present disclosure are administered in combination with one or more second agent(s) selected from a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a cytokine, an A2A antagonist, a GITR agonist, a TIM-3 inhibitor, a STING agonist, and a TLR7 agonist, to treat a disease, e.g., cancer.

In another embodiment, one or more chemotherapeutic agents are used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer, wherein said chemotherapeutic agents include, but are not limited to, anastrozole (Arimidex®), bicalutamide (Casodex®), bleomycin sulfate (Blenoxane®), busulfan (Myleran®), busulfan injection (Busulfex®), capecitabine (Xeloda®), N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin (Paraplatin®), carmustine (BiCNU®), chlorambucil (Leukeran®), cisplatin (Platinol®), cladribine (Leustatin®), cyclophosphamide (Cytoxan or Neosar®), cytarabine, cytosine arabinoside (Cytosar-U®), cytarabine liposome injection (DepoCyt®), dacarbazine (DTIC-Dome®), dactinomycin (Actinomycin D, Cosmegan), daunorubicin hydrochloride (Cerubidine®), daunorubicin citrate liposome injection (DaunoXome®), dexamethasone, docetaxel (Taxotere®), doxorubicin hydrochloride (Adriamycin®, Rubex®), etoposide (Vepesid®), fludarabine phosphate (Fludara®), 5-fluorouracil (Adrucil®, Efudex®), flutamide (Eulexin®), tezacitibine, Gemcitabine (difluorodeoxycitidine), hydroxyurea (Hydrea®), Idarubicin (Idamycin®), ifosfamide (IFEX®), irinotecan (Camptosar®), L-asparaginase (ELSPAR®), leucovorin calcium, melphalan (Alkeran®), 6-mercaptopurine (Purinethol®), methotrexate (Folex®), mitoxantrone (Novantrone®), mylotarg, paclitaxel (Taxol®), phoenix (Yttrium90/MX-DTPA), pentostatin, polifeprosan 20 with carmustine implant (Gliadel®), tamoxifen citrate (Nolvadex®), teniposide (Vumon®), 6-thioguanine, thiotepa, tirapazamine (Tirazone®), topotecan hydrochloride for injection (Hycamptin®), vinblastine (Velban®), vincristine (Oncovin®), vinorelbine (Navelbine®), epirubicin (Ellence®), oxaliplatin (Eloxatin®), exemestane (Aromasin®), letrozole (Femara®), and fulvestrant (Faslodex®).

In other embodiments, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more other anti-HER2 antibodies, e.g., trastuzumab, pertuzumab, margetuximab, or HT-19 described above, or with other anti-HER2 conjugates, e.g., ado-trastuzumab emtansine (also known as Kadcyla®, or T-DM1).

In other embodiments, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more tyrosine kinase inhibitors, including but not limited to, EGFR inhibitors, Her3 inhibitors, IGFR inhibitors, and Met inhibitors, for treating a disease, e.g., cancer.

For example, tyrosine kinase inhibitors include but are not limited to, Erlotinib hydrochloride (Tarceva®); Linifanib (N-[4-(3-amino-1H-indazol-4-yl)phenyl]-N′-(2-fluoro-5-methylphenyl)urea, also known as ABT 869, available from Genentech); Sunitinib malate (Sutent®); Bosutinib (4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methylpiperazin-1-yl)propoxy]quinoline-3-carbonitrile, also known as SKI-606, and described in U.S. Pat. No. 6,780,996); Dasatinib (Sprycel®); Pazopanib (Votrient®); Sorafenib (Nexavar®); Zactima (ZD6474); and Imatinib or Imatinib mesylate (Gilvec® and Gleevec®).

Epidermal growth factor receptor (EGFR) inhibitors include but are not limited to, Erlotinib hydrochloride (Tarceva®), Gefitinib (Iressa®); N-[4-[(3-Chloro-4-fluorophenyl)amino]-7-[[(3“S”)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4(dimethylamino)-2-butenamide, Tovok®); Vandetanib (Caprelsa®); Lapatinib (Tykerb®); (3R,4R)-4-Amino-1-((4-((3-methoxyphenyl)amino)pyrrolo[2,1-f][1,2,4]triazin-5-yl)methyl)piperidin-3-ol (BMS690514); Canertinib dihydrochloride (CI-1033); 6-[4-[(4-Ethyl-1-piperazinyl)methyl]phenyl]-N-[(1R)-1-phenylethyl]-7H-Pyrrolo[2,3-d]pyrimidin-4-amine (AEE788, CAS 497839-62-0); Mubritinib (TAK165); Pelitinib (EKB569); Afatinib (Gilotrif®); Neratinib (HKI-272); N-[4-[[1-[(3-Fluorophenyl)methyl]-1H-indazol-5-yl]amino]-5-methylpyrrolo[2,1-f][1,2,4]triazin-6-yl]-carbamic acid, (3S)-3-morpholinylmethyl ester (BMS599626); N-(3,4-Dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aα,5β,6aα)-octahydro-2-methylcyclopenta[c]pyrrol-5-yl]methoxy]-4-quinazolinamine (XL647, CAS 781613-23-8); and 4-[4-[[(1R)-1-Phenylethyl]amino]-7H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol (PKI166, CAS187724-61-4).

EGFR antibodies include but are not limited to, Cetuximab (Erbitux®); Panitumumab (Vectibix®); Matuzumab (EMD-72000); Nimotuzumab (hR3); Zalutumumab; TheraCIM h-R3; MDX0447 (CAS 339151-96-1); and ch806 (mAb-806, CAS 946414-09-1).

Other HER2 inhibitors include but are not limited to, Neratinib (HKI-272, (2E)-N-[4-[[3-chloro-4-[(pyridin-2-yl)methoxy]phenyl]amino]-3-cyano-7-ethoxyquinolin-6-yl]-4-(dimethylamino)but-2-enamide, and described PCT Publication No. WO 05/028443); Lapatinib or Lapatinib ditosylate (Tykerb®); (3R,4R)-4-amino-1-((4-((3-methoxyphenyl)amino)pyrrolo[2,1-f][1,2,4]triazin-5-yl)methyl)piperidin-3-ol (BMS690514); (2E)-N-[4-[(3-Chloro-4-fluorophenyl)amino]-7-[[(3S)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4-(dimethylamino)-2-butenamide (BIBW-2992, CAS 850140-72-6); N-[4-[[1-[(3-Fluorophenyl)methyl]-1H-indazol-5-yl]amino]-5-methylpyrrolo[2,1-f][1,2,4]triazin-6-yl]-carbamic acid, (3S)-3-morpholinylmethyl ester (BMS 599626, CAS 714971-09-2); Canertinib dihydrochloride (PD183805 or CI-1033); and N-(3,4-Dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aα,5β,6aα)-octahydro-2-methylcyclopenta[c]pyrrol-5-yl]methoxy]-4-quinazolinamine (XL647, CAS 781613-23-8).

HER3 inhibitors include but are not limited to, LJM716, MM-121, AMG-888, RG7116, REGN-1400, AV-203, MP-RM-1, MM-111, and MEHD-7945A.

MET inhibitors include but are not limited to, Cabozantinib (XL184, CAS 849217-68-1); Foretinib (GSK1363089, formerly XL880, CAS 849217-64-7); Tivantinib (ARQ197, CAS 1000873-98-2); 1-(2-Hydroxy-2-methylpropyl)-N-(5-(7-methoxy quinolin-4-yloxy)pyridin-2-yl)-5-methyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazole-4-carboxamide (AMG 458); Cryzotinib (Xalkori®, PF-02341066); (3Z)-5-(2,3-Dihydro-1H-indol-1-ylsulfonyl)-3-({3,5-dimethyl-4-[(4-methylpiperazin-1-yl)carbonyl]-1H-pyrrol-2-yl}methylene)-1,3-dihydro-2H-indol-2-one (SU11271); (3Z)—N-(3-Chlorophenyl)-3-({3,5-dimethyl-4-[(4-methylpiperazin-1-yl)carbonyl]-1H-pyrrol-2-yl}methylene)-N-methyl-2-oxoindoline-5-sulfonamide (SU11274); (3Z)—N-(3-Chlorophenyl)-3-{[3,5-dimethyl-4-(3-morpholin-4-ylpropyl)-1H-pyrrol-2-yl]methylene}-N-methyl-2-oxoindoline-5-sulfonamide (SU11606); 6-[Difluoro[6-(1-methyl-1Hpyrazol-4-yl)-1,2,4-triazolo[4,3-b]pyridazin-3-yl]methyl]-quinoline (JNJ38877605, CAS 943540-75-8); 2-[4-[1-(Quinolin-6-ylmethyl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-6-yl]-1H-pyrazol-1-yl]ethanol (PF04217903, CAS 956905-27-4); N-((2R)-1,4-Dioxan-2-ylmethyl)-N-methyl-N′-[3-(1-methyl-1H-pyrazol-4-yl)-5-oxo-5H-benzo[4,5]cyclohepta[1,2-b]pyridin-7-yl]sulfamide (MK2461, CAS 917879-39-1); 6-[[6-(1-Methyl-1H-pyrazol-4-yl)-1,2,4-triazolo[4,3-b]pyridazin 3-yl]thio]-quinoline (SGX523, CAS 1022150-57-7); and (3Z)-5-[[(2,6-Dichlorophenyl)methyl]sulfonyl]-3-[[3,5-dimethyl-4-[[(2R)-2-(1-pyrrolidinylmethyl)-1-pyrrolidinyl]carbonyl]-1H-pyrrol-2-yl]methylene]-1,3-dihydro-2H-indol-2-one (PHA665752, CAS 477575-56-7).

IGFR inhibitors include but are not limited to, BMS-754807, XL-228, OSI-906, GSK0904529A, A-928605, AXL1717, KW-2450, MK0646, AMG479, IMCA12, MEDI-573, and B1836845. See e.g., Yee, JNCI, 104; 975 (2012) for review.

In another embodiment, the compounds of Formula (I′) or Formula (I) of the present disclosure are used in combination with one or more proliferation signalling pathway inhibitors, including but not limited to, MEK inhibitors, BRAF inhibitors, PI3K/Akt inhibitors, SHP2 inhibitors, and also mTOR inhibitors, and CDK inhibitors, for treating a disease, e.g., cancer.

For example, mitogen-activated protein kinase (MEK) inhibitors include but are not limited to, XL-518 (also known as GDC-0973, CAS No. 1029872-29-4, available from ACC Corp.); 2-[(2-Chloro-4-iodophenyl)amino]-N-(cyclopropylmethoxy)-3,4-difluoro-benzamide (also known as CI-1040 or PD184352 and described in PCT Publication No. WO2000035436); N-[(2R)-2,3-Dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide (also known as PD0325901 and described in PCT Publication No. WO2002006213); 2,3-Bis[amino[(2-aminophenyl)thio]methylene]-butanedinitrile (also known as U0126 and described in U.S. Pat. No. 2,779,780); N-[3,4-Difluoro-2-[(2-fluoro-4-iodophenyl)amino]-6-methoxyphenyl]-1-[(2R)-2,3-dihydroxypropyl]-cyclopropanesulfonamide (also known as RDEA119 or BAY869766 and described in PCT Publication No. WO2007014011); (3S,4R,5Z,8S,9S,11E)-14-(Ethylamino)-8,9,16-trihydroxy-3,4-dimethyl-3,4,9, 19-tetrahydro-1H-2-benzoxacyclotetradecine-1,7(8H)-dione] (also known as E6201 and described in PCT Publication No. WO2003076424); 2′-Amino-3′-methoxyflavone (also known as PD98059 available from Biaffin GmbH & Co., KG, Germany); Vemurafenib (PLX-4032, CAS 918504-65-1); (R)-3-(2,3-Dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733, CAS 1035555-63-5); Pimasertib (AS-703026, CAS 1204531-26-9); and Trametinib dimethyl sulfoxide (GSK-1120212, CAS 1204531-25-80).

BRAF inhibitors include, but are not limited to, Vemurafenib (or Zelboraf®), GDC-0879, PLX-4720 (available from Symansis), Dabrafenib (or GSK2118436), LGX 818, CEP-32496, UI-152, RAF 265, Regorafenib (BAY 73-4506), CCT239065, or Sorafenib (or Sorafenib Tosylate, or Nexavar®), or Ipilimumab (or MDX-010, MDX-101, or Yervoy).

Phosphoinositide 3-kinase (PI3K) inhibitors include, but are not limited to, 4-[2-(1H-Indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (also known as GDC0941, RG7321, GNE0941, Pictrelisib, or Pictilisib; and described in PCT Publication Nos. WO 09/036082 and WO 09/055730); Tozasertib (VX680 or MK-0457, CAS 639089-54-6); (5Z)-5-[[4-(4-Pyridinyl)-6-quinolinyl]methylene]-2,4-thiazolidinedione (GSK1059615, CAS 958852-01-2); (1E,4S,4aR,5R,6aS,9aR)-5-(Acetyloxy)-1-[(di-2-propenylamino)methylene]-4,4a,5,6,6a,8,9,9a-octahydro-11-hydroxy-4-(methoxymethyl)-4a,6a-dimethylcyclopenta[5,6]naphtho[1,2-c]pyran-2,7,10(1H)-trione (PX866, CAS 502632-66-8); 8-Phenyl-2-(morpholin-4-yl)-chromen-4-one (LY294002, CAS 154447-36-6); (S)—N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2-yl)pyrrolidine-1,2-dicarboxamide (also known as BYL719 or Alpelisib); 2-(4-(2-(1-isopropyl-3-methyl-1H-1,2,4-triazol-5-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepin-9-yl)-1H-pyrazol-1-yl)-2-methylpropanamide (also known as GDC0032, RG7604, or Taselisib).

mTOR inhibitors include but are not limited to, Temsirolimus (Torisel®); Ridaforolimus (formally known as deferolimus, (1R,2R,4S)-4-[(2R)-2 [(1R,9S,12S,15R,16E,18R,19R,21R,23S,24E,26E,28Z,30S,32S,35R)-1,18-dihydroxy-19,30-dimethoxy-15,17,21,23, 29,35-hexamethyl-2,3,10,14,20-pentaoxo-11,36-dioxa-4-azatricyclo[30.3.1.04,9]hexatriaconta-16,24,26,28-tetraen-12-yl]propyl]-2-methoxycyclohexyl dimethylphosphinate, also known as AP23573 and MK8669, and described in PCT Publication No. WO 03/064383); Everolimus (Afinitor® or RAD001); Rapamycin (AY22989, Sirolimus®); Simapimod (CAS 164301-51-3); (5-{2,4-Bis[(3S)-3-methylmorpholin-4-yl]pyrido[2,3-d]pyrimidin-7-yl}-2-methoxyphenyl)methanol (AZD8055); 2-Amino-8-[trans-4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxy-3-pyridinyl)-4-methyl-pyrido[2,3-d]pyrimidin-7(8H)-one (PF04691502, CAS 1013101-36-4); and N²-[1,4-dioxo-4-[[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholinium-4-yl]methoxy]butyl]-L-arginylglycyl-L-□-aspartylL-serine-, inner salt (SF1126, CAS 936487-67-1).

CDK inhibitors include but are not limited to, Palbociclib (also known as PD-0332991, Ibrance®, 6-Acetyl-8-cyclopentyl-5-methyl-2-{[5-(1-piperazinyl)-2-pyridinyl]amino}pyrido[2,3-d]pyrimidin-7(8H)-one).

In yet another embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more pro-apoptotics, including but not limited to, IAP inhibitors, BCL2 inhibitors, MCL1 inhibitors, TRAIL agents, CHK inhibitors, for treating a disease, e.g., cancer.

For examples, IAP inhibitors include but are not limited to, LCL161, GDC-0917, AEG-35156, AT406, and TL32711. Other examples of IAP inhibitors include but are not limited to those disclosed in WO04/005284, WO 04/007529, WO05/097791, WO 05/069894, WO 05/069888, WO 05/094818, US2006/0014700, US2006/0025347, WO 06/069063, WO 06/010118, WO 06/017295, and WO08/134679, all of which are incorporated herein by reference.

BCL-2 inhibitors include but are not limited to, 4-[4-[[2-(4-Chlorophenyl)-5,5-dimethyl-1-cyclohexen-1-yl]methyl]-1-piperazinyl]-N-[[4-[[(1R)-3-(4-morpholinyl)-1-[(phenylthio)methyl]propyl]amino]-3-[(trifluoromethyl)sulfonyl]phenyl]sulfonyl]benzamide (also known as ABT-263 and described in PCT Publication No. WO 09/155386); Tetrocarcin A; Antimycin; Gossypol ((−)BL-193); Obatoclax; Ethyl-2-amino-6-cyclopentyl-4-(1-cyano-2-ethoxy-2-oxoethyl)-4Hchromone-3-carboxylate (HA14-1); Oblimersen (G3139, Genasense®); Bak BH3 peptide; (−)-Gossypol acetic acid (AT-101); 4-[4-[(4′-Chloro[1,1′-biphenyl]-2-yl)methyl]-1-piperazinyl]-N-[[4-[[(1R)-3-(dimethylamino)-1-[(phenylthio)methyl]propyl]amino]-3-nitrophenyl]sulfonyl]-benzamide (ABT-737, CAS 852808-04-9); and Navitoclax (ABT-263, CAS 923564-51-6).

Proapoptotic receptor agonists (PARAs) including DR4 (TRAILR1) and DR5 (TRAILR2), including but are not limited to, Dulanermin (AMG-951, RhApo2L/TRAIL); Mapatumumab (HRS-ETR1, CAS 658052-09-6); Lexatumumab (HGS-ETR2, CAS 845816-02-6); Apomab (Apomab®); Conatumumab (AMG655, CAS 896731-82-1); and Tigatuzumab(CS1008, CAS 946415-34-5, available from Daiichi Sankyo).

Checkpoint Kinase (CHK) inhibitors include but are not limited to, 7-Hydroxystaurosporine (UCN-01); 6-Bromo-3-(1-methyl-1H-pyrazol-4-yl)-5-(3R)-3-piperidinylpyrazolo[1,5-a]pyrimidin-7-amine (SCH900776, CAS 891494-63-6); 5-(3-Fluorophenyl)-3-ureidothiophene-2-carboxylic acid N—[(S)-piperidin-3-yl]amide (AZD7762, CAS 860352-01-8); 4-[((3S)-1-Azabicyclo[2.2.2]oct-3-yl)amino]-3-(1H-benzimidazol-2-yl)-6-chloroquinolin-2(1H)-one (CHIR 124, CAS 405168-58-3); 7-Aminodactinomycin (7-AAD), Isogranulatimide, debromohymenialdisine; N-[5-Bromo-4-methyl-2-[(2S)-2-morpholinylmethoxy]-phenyl]-N′-(5-methyl-2-pyrazinyl)urea (LY2603618, CAS 911222-45-2); Sulforaphane (CAS 4478-93-7, 4-Methylsulfinylbutyl isothiocyanate); 9,10,11,12-Tetrahydro-9,12-epoxy-1H-diindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocine-1,3(2H)-dione (SB-218078, CAS 135897-06-2); and TAT-S216A (YGRKKRRQRRRLYRSPAMPENL (SEQ ID NO: 33)), and CBP501 ((d-Bpa)sws(d-Phe-F5)(d-Cha)rrrqrr).

In a further embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more immunomodulators (e.g., one or more of an activator of a costimulatory molecule or an inhibitor of an immune checkpoint molecule), for treating a disease, e.g., cancer.

In certain embodiments, the immunomodulator is an activator of a costimulatory molecule. In one embodiment, the agonist of the costimulatory molecule is selected from an agonist (e.g., an agonistic antibody or antigen-binding fragment thereof, or a soluble fusion) of OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3 or CD83 ligand.

GITR Agonists

In some embodiments, a GITR agonist is used in combination with a compound of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the GITR agonist is GWN323 (Novartis), BMS-986156, MK-4166 or MK-1248 (Merck), TRX518 (Leap Therapeutics), INCAGN1876 (Incyte/Agenus), AMG 228 (Amgen) or INBRX-110 (Inhibrx).

Exemplary GITR Agonists

In one embodiment, the GITR agonist is an anti-GITR antibody molecule. In one embodiment, the GITR agonist is an anti-GITR antibody molecule as described in WO 2016/057846, published on Apr. 14, 2016, entitled “Compositions and Methods of Use for Augmented Immune Response and Cancer Therapy,” incorporated by reference in its entirety.

In one embodiment, the anti-GITR antibody molecule comprises at least one, two, three, four, five or six complementarity determining regions (CDRs) (or collectively all of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 1 (e.g., from the heavy and light chain variable region sequences of MAB7 disclosed in Table 1), or encoded by a nucleotide sequence shown in Table 1. In some embodiments, the CDRs are according to the Kabat definition (e.g., as set out in Table 1). In some embodiments, the CDRs are according to the Chothia definition (e.g., as set out in Table 1). In one embodiment, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions (e.g., conservative amino acid substitutions) or deletions, relative to an amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1.

In one embodiment, the anti-GITR antibody molecule comprises a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence of SEQ ID NO: 9, a VHCDR2 amino acid sequence of SEQ ID NO: 11, and a VHCDR3 amino acid sequence of SEQ ID NO: 13; and a light chain variable region (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 14, a VLCDR2 amino acid sequence of SEQ ID NO: 16, and a VLCDR3 amino acid sequence of SEQ ID NO: 18, each disclosed in Table 1.

In one embodiment, the anti-GITR antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 1. In one embodiment, the anti-GITR antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 2, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 2. In one embodiment, the anti-GITR antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 1 and a VL comprising the amino acid sequence of SEQ ID NO: 2.

In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 5, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 5. In one embodiment, the antibody molecule comprises a VL encoded by the nucleotide sequence of SEQ ID NO: 6, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 6. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 5 and a VL encoded by the nucleotide sequence of SEQ ID NO: 6.

In one embodiment, the anti-GITR antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 3, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 3. In one embodiment, the anti-GITR antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 4, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 4. In one embodiment, the anti-GITR antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 3 and a light chain comprising the amino acid sequence of SEQ ID NO: 4.

In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 7, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 7. In one embodiment, the antibody molecule comprises a light chain encoded by the nucleotide sequence of SEQ ID NO: 8, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 8. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 7 and a light chain encoded by the nucleotide sequence of SEQ ID NO: 8.

The antibody molecules described herein can be made by vectors, host cells, and methods described in WO 2016/057846, incorporated by reference in its entirety.

TABLE 1 Amino acid and nucleotide sequences of exemplary anti-GITR antibody molecule MAB7 SEQ ID NO: 1 VH EVQLVESGGGLVQSGGSLRLSCAASGFSLSSYGVDWVRQAPGKG LEWVGVIWGGGGTYYASSLMGRFTISRDNSKNTLYLQMNSLRAE DTAVYYCARHAYGHDGGFAMDYWGQGTLVTVSS SEQ ID NO: 2 VL EIVMTQSPATLSVSPGERATLSCRASESVSSNVAWYQQRPGQAPR LLIYGASNRATGIPARFSGSGSGTDFTLTISRLEPEDFAVYYCGQSY SYPFTFGQGTKLEIK SEQ ID NO: 3 Heavy EVQLVESGGGLVQSGGSLRLSCAASGFSLSSYGVDWVRQAPGKG Chain LEWVGVIWGGGGTYYASSLMGRFTISRDNSKNTLYLQMNSLRAE DTAVYYCARHAYGHDGGFAMDYWGQGTLVTVSSASTKGPSVFP LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEP KSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK SLSLSPGK SEQ ID NO: 4 Light EIVMTQSPATLSVSPGERATLSCRASESVSSNVAWYQQRPGQAPR Chain LLIYGASNRATGIPARFSGSGSGTDFTLTISRLEPEDFAVYYCGQSY SYPFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 5 DNA GAGGTGCAGCTGGTGGAATCTGGCGGCGGACTGGTGCAGTCCG VH GCGGCTCTCTGAGACTGTCTTGCGCTGCCTCCGGCTTCTCCCTG TCCTCTTACGGCGTGGACTGGGTGCGACAGGCCCCTGGCAAGG GCCTGGAATGGGTGGGAGTGATCTGGGGCGGAGGCGGCACCT ACTACGCCTCTTCCCTGATGGGCCGGTTCACCATCTCCCGGGAC AACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGCGGG CCGAGGACACCGCCGTGTACTACTGCGCCAGACACGCCTACGG CCACGACGGCGGCTTCGCCATGGATTATTGGGGCCAGGGCACC CTGGTGACAGTGTCCTCC SEQ ID NO: 6 DNA GAGATCGTGATGACCCAGTCCCCCGCCACCCTGTCTGTGTCTCC VL CGGCGAGAGAGCCACCCTGAGCTGCAGAGCCTCCGAGTCCGTG TCCTCCAACGTGGCCTGGTATCAGCAGAGACCTGGTCAGGCCC CTCGGCTGCTGATCTACGGCGCCTCTAACCGGGCCACCGGCAT CCCTGCCAGATTCTCCGGCTCCGGCAGCGGCACCGACTTCACC CTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACT ACTGCGGCCAGTCCTACTCATACCCCTTCACCTTCGGCCAGGG CACCAAGCTGGAAATCAAG SEQ ID NO: 7 DNA GAGGTGCAGCTGGTGGAATCTGGCGGCGGACTGGTGCAGTCCG Heavy GCGGCTCTCTGAGACTGTCTTGCGCTGCCTCCGGCTTCTCCCTG Chain TCCTCTTACGGCGTGGACTGGGTGCGACAGGCCCCTGGCAAGG GCCTGGAATGGGTGGGAGTGATCTGGGGCGGAGGCGGCACCT ACTACGCCTCTTCCCTGATGGGCCGGTTCACCATCTCCCGGGAC AACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGCGGG CCGAGGACACCGCCGTGTACTACTGCGCCAGACACGCCTACGG CCACGACGGCGGCTTCGCCATGGATTATTGGGGCCAGGGCACC CTGGTGACAGTGTCCTCCGCTAGCACCAAGGGCCCAAGTGTGT TTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGC TGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTG ACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACA CCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAG CAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACC TATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGG ACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCT GCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGT GTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGC AGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGTCCCACG AGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGG AGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACA ACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCA GGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAA CAAGGCCCTGCCAGCCCCAATCGAAAAGACAATCAGCAAGGC CAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCC AGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGT CTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGG AGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCC CAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCT GACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAG CTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAG AAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 8 DNA GAGATCGTGATGACCCAGTCCCCCGCCACCCTGTCTGTGTCTCC Light CGGCGAGAGAGCCACCCTGAGCTGCAGAGCCTCCGAGTCCGTG Chain TCCTCCAACGTGGCCTGGTATCAGCAGAGACCTGGTCAGGCCC CTCGGCTGCTGATCTACGGCGCCTCTAACCGGGCCACCGGCAT CCCTGCCAGATTCTCCGGCTCCGGCAGCGGCACCGACTTCACC CTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACT ACTGCGGCCAGTCCTACTCATACCCCTTCACCTTCGGCCAGGG CACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTG TTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCG CCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGC CAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAA CAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCAC CTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTAC GAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGC CTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC SEQ ID NO: 9 HCDR1 SYGVD (KABAT) SEQ ID NO: 10 HCDR1 GFSLSSY (CHOTHIA) SEQ ID NO: 11 HCDR2 VIWGGGGTYYASSLMG (KABAT) SEQ ID NO: 12 HCDR2 WGGGG (CHOTHIA) SEQ ID NO: 13 HCDR3 HAYGHDGGFAMDY (KABAT) SEQ ID NO: 13 HCDR3 HAYGHDGGFAMDY (CHOTHIA) SEQ ID NO: 14 LCDR1 RASESVSSNVA (KABAT) SEQ ID NO: 15 LCDR1 SESVSSN (CHOTHIA) SEQ ID NO: 16 LCDR2 GASNRAT (KABAT) SEQ ID NO: 17 LCDR2 GAS (CHOTHIA) SEQ ID NO: 18 LCDR3 GQSYSYPFT (KABAT) SEQ ID NO: 19 LCDR3 SYSYPF (CHOTHIA) Other Exemplary GITR Agonists

In one embodiment, the anti-GITR antibody molecule is BMS-986156 (Bristol-Myers Squibb), also known as BMS 986156 or BMS986156. BMS-986156 and other anti-GITR antibodies are disclosed, e.g., in U.S. Pat. No. 9,228,016 and WO 2016/196792, incorporated by reference in their entirety. In one embodiment, the anti-GITR antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of BMS-986156, e.g., as disclosed in Table 2.

In one embodiment, the anti-GITR antibody molecule is MK-4166 or MK-1248 (Merck). MK-4166, MK-1248, and other anti-GITR antibodies are disclosed, e.g., in U.S. Pat. No. 8,709,424, WO 2011/028683, WO 2015/026684, and Mahne et al. Cancer Res. 2017; 77(5): 1108-1118, incorporated by reference in their entirety. In one embodiment, the anti-GITR antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of MK-4166 or MK-1248.

In one embodiment, the anti-GITR antibody molecule is TRX518 (Leap Therapeutics). TRX518 and other anti-GITR antibodies are disclosed, e.g., in U.S. Pat. Nos. 7,812,135, 8,388,967, 9,028,823, WO 2006/105021, and Ponte J et al. (2010) Clinical Immunology; 135:S96, incorporated by reference in their entirety. In one embodiment, the anti-GITR antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of TRX518.

In one embodiment, the anti-GITR antibody molecule is INCAGN1876 (Incyte/Agenus). INCAGN1876 and other anti-GITR antibodies are disclosed, e.g., in US 2015/0368349 and WO 2015/184099, incorporated by reference in their entirety. In one embodiment, the anti-GITR antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of INCAGN1876.

In one embodiment, the anti-GITR antibody molecule is AMG 228 (Amgen). AMG 228 and other anti-GITR antibodies are disclosed, e.g., in U.S. Pat. No. 9,464,139 and WO 2015/031667, incorporated by reference in their entirety. In one embodiment, the anti-GITR antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of AMG 228.

In one embodiment, the anti-GITR antibody molecule is INBRX-110 (Inhibrx). INBRX-110 and other anti-GITR antibodies are disclosed, e.g., in US 2017/0022284 and WO 2017/015623, incorporated by reference in their entirety. In one embodiment, the GITR agonist comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of INBRX-110.

In one embodiment, the GITR agonist (e.g., a fusion protein) is MEDI 1873 (MedImmune), also known as MEDI1873. MEDI 1873 and other GITR agonists are disclosed, e.g., in US 2017/0073386, WO 2017/025610, and Ross et al. Cancer Res 2016; 76(14 Suppl): Abstract nr 561, incorporated by reference in their entirety. In one embodiment, the GITR agonist comprises one or more of an IgG Fc domain, a functional multimerization domain, and a receptor binding domain of a glucocorticoid-induced TNF receptor ligand (GITRL) of MEDI 1873.

Further known GITR agonists (e.g., anti-GITR antibodies) include those described, e.g., in WO 2016/054638, incorporated by reference in its entirety.

In one embodiment, the anti-GITR antibody is an antibody that competes for binding with, and/or binds to the same epitope on GITR as, one of the anti-GITR antibodies described herein.

In one embodiment, the GITR agonist is a peptide that activates the GITR signalling pathway. In one embodiment, the GITR agonist is an immunoadhesin binding fragment (e.g., an immunoadhesin binding fragment comprising an extracellular or GITR binding portion of GITRL) fused to a constant region (e.g., an Fc region of an immunoglobulin sequence).

TABLE 2 Amino acid sequence of other exemplary anti-GITR antibody molecules BMS-986156 SEQ ID NO: 20 VH QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKG LEWVAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAE DTAVYYCARGGSMVRGDYYYGMDVWGQGTTVTVSS SEQ ID NO: 21 VL AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLL IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYP YTFGQGTKLEIK

In certain embodiments, the immunomodulator is an inhibitor of an immune checkpoint molecule. In one embodiment, the immunomodulator is an inhibitor of PD-1, PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGFRbeta. In one embodiment, the inhibitor of an immune checkpoint molecule inhibits PD-1, PD-L1, LAG-3, TIM-3 or CTLA4, or any combination thereof. The term “inhibition” or “inhibitor” includes a reduction in a certain parameter, e.g., an activity, of a given molecule, e.g., an immune checkpoint inhibitor. For example, inhibition of an activity, e.g., a PD-1 or PD-L1 activity, of at least 5%, 10%, 20%, 30%, 40%, 50% or more is included by this term. Thus, inhibition need not be 100%.

Inhibition of an inhibitory molecule can be performed at the DNA, RNA or protein level. In some embodiments, an inhibitory nucleic acid (e.g., a dsRNA, siRNA or shRNA), can be used to inhibit expression of an inhibitory molecule. In other embodiments, the inhibitor of an inhibitory signal is a polypeptide e.g., a soluble ligand (e.g., PD-1-Ig or CTLA-4 Ig), or an antibody or antigen-binding fragment thereof, that binds to the inhibitory molecule; e.g., an antibody or fragment thereof (also referred to herein as “an antibody molecule”) that binds to PD-1, PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGFR beta, or a combination thereof.

In one embodiment, the antibody molecule is a full antibody or fragment thereof (e.g., a Fab, F(ab′)2, Fv, or a single chain Fv fragment (scFv)). In yet other embodiments, the antibody molecule has a heavy chain constant region (Fc) selected from, e.g., the heavy chain constant regions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE; particularly, selected from, e.g., the heavy chain constant regions of IgG1, IgG2, IgG3, and IgG4, more particularly, the heavy chain constant region of IgG1 or IgG4 (e.g., human IgG1 or IgG4). In one embodiment, the heavy chain constant region is human IgG1 or human IgG4. In one embodiment, the constant region is altered, e.g., mutated, to modify the properties of the antibody molecule (e.g., to increase or decrease one or more of Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, or complement function).

In certain embodiments, the antibody molecule is in the form of a bispecific or multispecific antibody molecule. In one embodiment, the bispecific antibody molecule has a first binding specificity to PD-1 or PD-L1 and a second binding specificity, e.g., a second binding specificity to TIM-3, LAG-3, or PD-L2. In one embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and TIM-3. In another embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and LAG-3. In another embodiment, the bispecific antibody molecule binds to PD-1 and PD-L1. In yet another embodiment, the bispecific antibody molecule binds to PD-1 and PD-L2. In another embodiment, the bispecific antibody molecule binds to TIM-3 and LAG-3. Any combination of the aforesaid molecules can be made in a multispecific antibody molecule, e.g., a trispecific antibody that includes a first binding specificity to PD-1 or PD-1, and a second and third binding specificities to two or more of TIM-3, LAG-3, or PD-L2.

In certain embodiments, the immunomodulator is an inhibitor of PD-1, e.g., human PD-1. In another embodiment, the immunomodulator is an inhibitor of PD-L1, e.g., human PD-L1. In one embodiment, the inhibitor of PD-1 or PD-L1 is an antibody molecule to PD-1 or PD-L1. The PD-1 or PD-L1 inhibitor can be administered alone, or in combination with other immunomodulators, e.g., in combination with an inhibitor of LAG-3, TIM-3 or CTLA4. In an exemplary embodiment, the inhibitor of PD-1 or PD-L1, e.g., the anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a LAG-3 inhibitor, e.g., an anti-LAG-3 antibody molecule. In another embodiment, the inhibitor of PD-1 or PD-L1, e.g., the anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a TIM-3 inhibitor, e.g., an anti-TIM-3 antibody molecule. In yet other embodiments, the inhibitor of PD-1 or PD-L1, e.g., the anti-PD-1 antibody molecule, is administered in combination with a LAG-3 inhibitor, e.g., an anti-LAG-3 antibody molecule, and a TIM-3 inhibitor, e.g., an anti-TIM-3 antibody molecule.

Other combinations of immunomodulators with a PD-1 inhibitor (e.g., one or more of PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGFR) are also within the present disclosure. Any of the antibody molecules known in the art or disclosed herein can be used in the aforesaid combinations of inhibitors of checkpoint molecule.

PD-1 Inhibitors

In some embodiments, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with a PD-1 inhibitor to treat a disease, e.g., cancer. In some embodiments, the PD-1 inhibitor is selected from PDR001 (Novartis), Nivolumab (Bristol-Myers Squibb), Pembrolizumab (Merck & Co), Pidilizumab (CureTech), MEDI0680 (Medimmune), REGN2810 (Regeneron), TSR-042 (Tesaro), PF-06801591 (Pfizer), BGB-A317 (Beigene), BGB-108 (Beigene), INCSHR1210 (Incyte), or AMP-224 (Amplimmune).

Exemplary PD-1 Inhibitors

In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody molecule. In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody molecule as described in US 2015/0210769, published on Jul. 30, 2015, entitled “Antibody Molecules to PD-1 and Uses Thereof,” incorporated by reference in its entirety.

In one embodiment, the anti-PD-1 antibody molecule comprises at least one, two, three, four, five or six complementarity determining regions (CDRs) (or collectively all of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 3 (e.g., from the heavy and light chain variable region sequences of BAP049-Clone-E or BAP049-Clone-B disclosed in Table 3), or encoded by a nucleotide sequence shown in Table 3. In some embodiments, the CDRs are according to the Kabat definition (e.g., as set out in Table 3). In some embodiments, the CDRs are according to the Chothia definition (e.g., as set out in Table 3). In some embodiments, the CDRs are according to the combined CDR definitions of both Kabat and Chothia (e.g., as set out in Table 3). In one embodiment, the combination of Kabat and Chothia CDR of VH CDR1 comprises the amino acid sequence GYTFTTYWMH (SEQ ID NO: 213). In one embodiment, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions (e.g., conservative amino acid substitutions) or deletions, relative to an amino acid sequence shown in Table 3, or encoded by a nucleotide sequence shown in Table 3.

In one embodiment, the anti-PD-1 antibody molecule comprises a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence of SEQ ID NO: 22, a VHCDR2 amino acid sequence of SEQ ID NO: 23, and a VHCDR3 amino acid sequence of SEQ ID NO: 24; and a light chain variable region (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 31, a VLCDR2 amino acid sequence of SEQ ID NO: 32, and a VLCDR3 amino acid sequence of SEQ ID NO: 286, each disclosed in Table 3.

In one embodiment, the antibody molecule comprises a VH comprising a VHCDR1 encoded by the nucleotide sequence of SEQ ID NO: 45, a VHCDR2 encoded by the nucleotide sequence of SEQ ID NO: 46, and a VHCDR3 encoded by the nucleotide sequence of SEQ ID NO: 47; and a VL comprising a VLCDR1 encoded by the nucleotide sequence of SEQ ID NO: 50, a VLCDR2 encoded by the nucleotide sequence of SEQ ID NO: 51, and a VLCDR3 encoded by the nucleotide sequence of SEQ ID NO: 52, each disclosed in Table 3.

In one embodiment, the anti-PD-1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 27, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 27. In one embodiment, the anti-PD-1 antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 41, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 41. In one embodiment, the anti-PD-1 antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 37, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 37. In one embodiment, the anti-PD-1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 27 and a VL comprising the amino acid sequence of SEQ ID NO: 41. In one embodiment, the anti-PD-1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 27 and a VL comprising the amino acid sequence of SEQ ID NO: 37.

In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 28, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 28. In one embodiment, the antibody molecule comprises a VL encoded by the nucleotide sequence of SEQ ID NO: 42 or 38, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 42 or 38. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 28 and a VL encoded by the nucleotide sequence of SEQ ID NO: 42 or 38.

In one embodiment, the anti-PD-1 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 29. In one embodiment, the anti-PD-1 antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 43, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 43. In one embodiment, the anti-PD-1 antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 39, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 39. In one embodiment, the anti-PD-1 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 43. In one embodiment, the anti-PD-1 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 39.

In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 30, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 30. In one embodiment, the antibody molecule comprises a light chain encoded by the nucleotide sequence of SEQ ID NO: 44 or 40, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 44 or 40. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 30 and a light chain encoded by the nucleotide sequence of SEQ ID NO: 44 or 40.

The antibody molecules described herein can be made by vectors, host cells, and methods described in US 2015/0210769, incorporated by reference in its entirety.

TABLE 3 Amino acid and nucleotide sequences of exemplary anti-PD-1 antibody molecules BAP049-Clone-B HC SEQ ID NO: 22 HCDR1 TYWMH (Kabat) SEQ ID NO: 23 HCDR2 NIYPGTGGSNFDEKFKN (Kabat) SEQ ID NO: 24 HCDR3 WTTGTGAY (Kabat) SEQ ID NO: 25 HCDR1 GYTFTTY (Chothia) SEQ ID NO: 26 HCDR2 YPGTGG (Chothia) SEQ ID NO: 24 HCDR3 WTTGTGAY (Chothia) SEQ ID NO: 27 VH EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQAT GQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYME LSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 28 DNA GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAG VH CCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCAGGCT ACACCTTCACTACCTACTGGATGCACTGGGTCCGCCAGGC TACCGGTCAAGGCCTCGAGTGGATGGGTAATATCTACCCC GGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAATA GAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTA TATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTC TACTACTGCACTAGGTGGACTACCGGCACAGGCGCCTACT GGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: 29 Heavy EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQAT chain GQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYME LSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGP SVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNT KVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISK AKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 30 DNA GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAG heavy CCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCAGGCT chain ACACCTTCACTACCTACTGGATGCACTGGGTCCGCCAGGC TACCGGTCAAGGCCTCGAGTGGATGGGTAATATCTACCCC GGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAATA GAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTA TATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTC TACTACTGCACTAGGTGGACTACCGGCACAGGCGCCTACT GGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCAC TAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGG AGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCA AGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAG CGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTG CTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCA CGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGC AACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGC GCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCC CGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCC CACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCC TGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGAT CCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGG TGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCA ACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCAT CAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTG TCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCT CGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATA CCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAG TCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGAT ATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAAC AACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGAT CCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAG ATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCAT GAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCC TCTCCCTGGGA BAP049-Clone-B LC SEQ ID NO: 31 LCDR1 KSSQSLLDSGNQKNFLT (Kabat) SEQ ID NO: 32 LCDR2 WASTRES (Kabat) SEQ ID NO: 286 LCDR3 QNDYSYPYT (Kabat) SEQ ID NO: 34 LCDR1 SQSLLDSGNQKNF (Chothia) SEQ ID NO: 35 LCDR2 WAS (Chothia) SEQ ID NO: 36 LCDR3 DYSYPY (Chothia) SEQ ID NO: 37 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQ QKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLQPE DIATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 38 DNA GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGA VL GCCCTGGCGAGCGGGCTACACTGAGCTGTAAATCTAGTCA GTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACC TGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGA TCTACTGGGCCTCTACTAGAGAATCAGGCGTGCCCTCTAG GTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACT ATCTCTAGCCTGCAGCCCGAGGATATCGCTACCTACTACTG TCAGAACGACTATAGCTACCCCTACACCTTCGGTCAAGGC ACTAAGGTCGAGATTAAG SEQ ID NO: 39 Light EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQ chain QKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLQPE DIATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC SEQ ID NO: 40 DNA GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGA light GCCCTGGCGAGCGGGCTACACTGAGCTGTAAATCTAGTCA chain GTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACC TGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGA TCTACTGGGCCTCTACTAGAGAATCAGGCGTGCCCTCTAG GTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACT ATCTCTAGCCTGCAGCCCGAGGATATCGCTACCTACTACTG TCAGAACGACTATAGCTACCCCTACACCTTCGGTCAAGGC ACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCG TGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGG CACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCC CGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTG CAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGAC AGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCC TGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTG CGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAG AGCTTCAACAGGGGCGAGTGC BAP049-Clone-E HC SEQ ID NO: 22 HCDR1 TYWMH (Kabat) SEQ ID NO: 23 HCDR2 NIYPGTGGSNFDEKFKN (Kabat) SEQ ID NO: 24 HCDR3 WTTGTGAY (Kabat) SEQ ID NO: 25 HCDR1 GYTFTTY (Chothia) SEQ ID NO: 26 HCDR2 YPGTGG (Chothia) SEQ ID NO: 24 HCDR3 WTTGTGAY (Chothia) SEQ ID NO: 27 VH EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQAT GQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYME LSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 28 DNA GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAG VH CCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCAGGCT ACACCTTCACTACCTACTGGATGCACTGGGTCCGCCAGGC TACCGGTCAAGGCCTCGAGTGGATGGGTAATATCTACCCC GGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAATA GAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTA TATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTC TACTACTGCACTAGGTGGACTACCGGCACAGGCGCCTACT GGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: 29 Heavy EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQAT chain GQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYME LSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGP SVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNT KVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISK AKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 30 DNA GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAG heavy CCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCAGGCT chain ACACCTTCACTACCTACTGGATGCACTGGGTCCGCCAGGC TACCGGTCAAGGCCTCGAGTGGATGGGTAATATCTACCCC GGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAATA GAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTA TATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTC TACTACTGCACTAGGTGGACTACCGGCACAGGCGCCTACT GGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCAC TAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGG AGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCA AGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAG CGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTG CTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCA CGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGC AACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGC GCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCC CGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCC CACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCC TGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGAT CCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGG TGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCA ACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCAT CAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTG TCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCT CGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATA CCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAG TCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGAT ATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAAC AACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGAT CCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAG ATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCAT GAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCC TCTCCCTGGGA BAP049-Clone-E LC SEQ ID NO: 31 LCDR1 KSSQSLLDSGNQKNFLT (Kabat) SEQ ID NO: 32 LCDR2 WASTRES (Kabat) SEQ ID NO: 286 LCDR3 QNDYSYPYT (Kabat) SEQ ID NO: 34 LCDR1 SQSLLDSGNQKNF (Chothia) SEQ ID NO: 35 LCDR2 WAS (Chothia) SEQ ID NO: 36 LCDR3 DYSYPY (Chothia) SEQ ID NO: 41 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQ QKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAE DAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 42 DNA GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGA VL GCCCTGGCGAGCGGGCTACACTGAGCTGTAAATCTAGTCA GTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACC TGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGA TCTACTGGGCCTCTACTAGAGAATCAGGCGTGCCCTCTAG GTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACT ATCTCTAGCCTGGAAGCCGAGGACGCCGCTACCTACTACT GTCAGAACGACTATAGCTACCCCTACACCTTCGGTCAAGG CACTAAGGTCGAGATTAAG SEQ ID NO: 43 Light EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQ chain QKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAE DAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC SEQ ID NO: 44 DNA GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGA light GCCCTGGCGAGCGGGCTACACTGAGCTGTAAATCTAGTCA chain GTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACC TGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGA TCTACTGGGCCTCTACTAGAGAATCAGGCGTGCCCTCTAG GTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACT ATCTCTAGCCTGGAAGCCGAGGACGCCGCTACCTACTACT GTCAGAACGACTATAGCTACCCCTACACCTTCGGTCAAGG CACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGC GTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCG GCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCC CCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCT GCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACC CTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCT GCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAA GAGCTTCAACAGGGGCGAGTGC BAP049-Clone-B HC SEQ ID NO: 45 HCDR1 ACCTACTGGATGCAC (Kabat) SEQ ID NO: 46 HCDR2 AATATCTACCCCGGCACCGGCGGCTCTAACTTCGACGAGA (Kabat) AGTTTAAGAAT SEQ ID NO: 47 HCDR3 TGGACTACCGGCACAGGCGCCTAC (Kabat) SEQ ID NO: 48 HCDR1 GGCTACACCTTCACTACCTAC (Chothia) SEQ ID NO: 49 HCDR2 TACCCCGGCACCGGCGGC (Chothia) SEQ ID NO: 47 HCDR3 TGGACTACCGGCACAGGCGCCTAC (Chothia) BAP049-Clone-B LC SEQ ID NO: 50 LCDR1 AAATCTAGTCAGTCACTGCTGGATAGCGGTAATCAGAAGA (Kabat) ACTTCCTGACC SEQ ID NO: 51 LCDR2 TGGGCCTCTACTAGAGAATCA (Kabat) SEQ ID NO: 52 LCDR3 CAGAACGACTATAGCTACCCCTACACC (Kabat) SEQ ID NO: 53 LCDR1 AGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTC (Chothia) SEQ ID NO: 54 LCDR2 TGGGCCTCT (Chothia) SEQ ID NO: 55 LCDR3 GACTATAGCTACCCCTAC (Chothia) BAP049-Clone-E HC SEQ ID NO: 45 HCDR1 ACCTACTGGATGCAC (Kabat) SEQ ID NO: 46 HCDR2 AATATCTACCCCGGCACCGGCGGCTCTAACTTCGACGAGA (Kabat) AGTTTAAGAAT SEQ ID NO: 47 HCDR3 TGGACTACCGGCACAGGCGCCTAC (Kabat) SEQ ID NO: 48 HCDR1 GGCTACACCTTCACTACCTAC (Chothia) SEQ ID NO: 49 HCDR2 TACCCCGGCACCGGCGGC (Chothia) SEQ ID NO: 47 HCDR3 TGGACTACCGGCACAGGCGCCTAC (Chothia) BAP049-Clone-E LC SEQ ID NO: 50 LCDR1 AAATCTAGTCAGTCACTGCTGGATAGCGGTAATCAGAAGA (Kabat) ACTTCCTGACC SEQ ID NO: 51 LCDR2 TGGGCCTCTACTAGAGAATCA (Kabat) SEQ ID NO: 52 LCDR3 CAGAACGACTATAGCTACCCCTACACC (Kabat) SEQ ID NO: 53 LCDR1 AGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTC (Chothia) SEQ ID NO: 54 LCDR2 TGGGCCTCT (Chothia) SEQ ID NO: 55 LCDR3 GACTATAGCTACCCCTAC (Chothia) Other Exemplary PD-1 Inhibitors

In some embodiments, the anti-PD-1 antibody is Nivolumab (CAS Registry Number: 946414-94-4). Alternative names for Nivolumab include MDX-1106, MDX-1106-04, ONO-4538, BMS-936558 or OPDIVO®. Nivolumab is a fully human IgG4 monoclonal antibody, which specifically blocks PD1. Nivolumab (clone 5C4) and other human monoclonal antibodies that specifically bind to PD1 are disclosed in U.S. Pat. No. 8,008,449 and PCT Publication No. WO2006/121168, incorporated by reference in their entirety. In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of Nivolumab, e.g., as disclosed in Table 4.

In other embodiments, the anti-PD-1 antibody is Pembrolizumab. Pembrolizumab (Trade name KEYTRUDA formerly Lambrolizumab, also known as Merck 3745, MK-3475 or SCH-900475) is a humanized IgG4 monoclonal antibody that binds to PD1. Pembrolizumab is disclosed, e.g., in Hamid, O. et al. (2013) New England Journal of Medicine 369 (2): 134-44, PCT Publication No. WO2009/114335, and U.S. Pat. No. 8,354,509, incorporated by reference in their entirety. In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of Pembrolizumab, e.g., as disclosed in Table 4.

In some embodiments, the anti-PD-1 antibody is Pidilizumab. Pidilizumab (CT-011; Cure Tech) is a humanized IgG1k monoclonal antibody that binds to PD1. Pidilizumab and other humanized anti-PD-1 monoclonal antibodies are disclosed in PCT Publication No. WO2009/101611, incorporated by reference in their entirety. In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of Pidilizumab, e.g., as disclosed in Table 4.

Other anti-PD1 antibodies are disclosed in U.S. Pat. No. 8,609,089, US Publication No. 2010028330, and/or US Publication No. 20120114649, incorporated by reference in their entirety. Other anti-PD1 antibodies include AMP 514 (Amplimmune).

In one embodiment, the anti-PD-1 antibody molecule is MEDI0680 (Medimmune), also known as AMP-514. MEDI0680 and other anti-PD-1 antibodies are disclosed in U.S. Pat. No. 9,205,148 and WO 2012/145493, incorporated by reference in their entirety. In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of MEDI0680.

In one embodiment, the anti-PD-1 antibody molecule is REGN2810 (Regeneron). In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of REGN2810.

In one embodiment, the anti-PD-1 antibody molecule is PF-06801591 (Pfizer). In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of PF-06801591.

In one embodiment, the anti-PD-1 antibody molecule is BGB-A317 or BGB-108 (Beigene). In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of BGB-A317 or BGB-108.

In one embodiment, the anti-PD-1 antibody molecule is INCSHR1210 (Incyte), also known as INCSHR01210 or SHR-1210. In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of INCSHR1210.

In one embodiment, the anti-PD-1 antibody molecule is TSR-042 (Tesaro), also known as ANB011. In one embodiment, the anti-PD-1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of TSR-042.

Further known anti-PD-1 antibodies include those described, e.g., in WO 2015/112800, WO 2016/092419, WO 2015/085847, WO 2014/179664, WO 2014/194302, WO 2014/209804, WO 2015/200119, U.S. Pat. Nos. 8,735,553, 7,488,802, 8,927,697, 8,993,731, and 9,102,727, incorporated by reference in their entirety.

In one embodiment, the anti-PD-1 antibody is an antibody that competes for binding with, and/or binds to the same epitope on PD-1 as, one of the anti-PD-1 antibodies described herein.

In one embodiment, the PD-1 inhibitor is a peptide that inhibits the PD-1 signalling pathway, e.g., as described in U.S. Pat. No. 8,907,053, incorporated by reference in its entirety. In some embodiments, the PD-1 inhibitor is an immunoadhesin (e.g., an immunoadhesin comprising an extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region (e.g., an Fc region of an immunoglobulin sequence). In some embodiments, the PD-1 inhibitor is AMP-224 (B7-DCIg (Amplimmune), e.g., disclosed in WO 2010/027827 and WO 2011/066342, incorporated by reference in their entirety).

TABLE 4 Amino acid sequences of other exemplary anti-PD-1 antibody molecules Nivolumab SEQ ID NO: 56 Heavy QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGL chain EWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAED TAVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTA ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV TVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV EVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKG LPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 57 Light EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI chain YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWP RTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC Pembrolizumab SEQ ID NO: 58 Heavy QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQG chain LEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDD TAVYYCARRDYRFDMGFDYWGQGTTVTVSSASTKGPSVFPLAPCS RSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPC PAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFN WYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDK SRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 59 Light EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQA chain PRLLIYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHS RDLPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC Pidilizumab SEQ ID NO: 60 Heavy QVQLVQSGSELKKPGASVKISCKASGYTFTNYGMNWVRQAPGQGL chain QWMGWINTDSGESTYAEEFKGRFVFSLDTSVNTAYLQITSLTAEDT GMYFCVRVGYDALDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCP APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 61 Light EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWFQQKPGKAPKLWI chain YRTSNLASGVPSRFSGSGSGTSYCLTINSLQPEDFATYYCQQRSSFPL TFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGEC PD-L1 Inhibitors

In some embodiments, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with a PD-L1 inhibitor for treating a disease, e.g., cancer. In some embodiments, the PD-L1 inhibitor is selected from FAZ053 (Novartis), Atezolizumab (Genentech/Roche), Avelumab (Merck Serono and Pfizer), Durvalumab (MedImmune/AstraZeneca), or BMS-936559 (Bristol-Myers Squibb).

Exemplary PD-L1 Inhibitors

In one embodiment, the PD-L1 inhibitor is an anti-PD-L1 antibody molecule. In one embodiment, the PD-L1 inhibitor is an anti-PD-L1 antibody molecule as disclosed in US 2016/0108123, published on Apr. 21, 2016, entitled “Antibody Molecules to PD-L1 and Uses Thereof,” incorporated by reference in its entirety.

In one embodiment, the anti-PD-L1 antibody molecule comprises at least one, two, three, four, five or six complementarity determining regions (CDRs) (or collectively all of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 5 (e.g., from the heavy and light chain variable region sequences of BAP058-Clone O or BAP058-Clone N disclosed in Table 5), or encoded by a nucleotide sequence shown in Table 5. In some embodiments, the CDRs are according to the Kabat definition (e.g., as set out in Table 5). In some embodiments, the CDRs are according to the Chothia definition (e.g., as set out in Table 5). In some embodiments, the CDRs are according to the combined CDR definitions of both Kabat and Chothia (e.g., as set out in Table 5). In one embodiment, the combination of Kabat and Chothia CDR of VH CDR1 comprises the amino acid sequence GYTFTSYWMY (SEQ ID NO: 214). In one embodiment, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions (e.g., conservative amino acid substitutions) or deletions, relative to an amino acid sequence shown in Table 5, or encoded by a nucleotide sequence shown in Table 5.

In one embodiment, the anti-PD-L1 antibody molecule comprises a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence of SEQ ID NO: 62, a VHCDR2 amino acid sequence of SEQ ID NO: 63, and a VHCDR3 amino acid sequence of SEQ ID NO: 64; and a light chain variable region (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 70, a VLCDR2 amino acid sequence of SEQ ID NO: 71, and a VLCDR3 amino acid sequence of SEQ ID NO: 72, each disclosed in Table 5.

In one embodiment, the anti-PD-L1 antibody molecule comprises a VH comprising a VHCDR1 encoded by the nucleotide sequence of SEQ ID NO: 89, a VHCDR2 encoded by the nucleotide sequence of SEQ ID NO: 90, and a VHCDR3 encoded by the nucleotide sequence of SEQ ID NO: 91; and a VL comprising a VLCDR1 encoded by the nucleotide sequence of SEQ ID NO: 94, a VLCDR2 encoded by the nucleotide sequence of SEQ ID NO: 95, and a VLCDR3 encoded by the nucleotide sequence of SEQ ID NO: 96, each disclosed in Table 5.

In one embodiment, the anti-PD-L1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 67, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 67. In one embodiment, the anti-PD-L1 antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 77, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 77. In one embodiment, the anti-PD-L1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 81, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 81. In one embodiment, the anti-PD-L1 antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 85, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 85. In one embodiment, the anti-PD-L1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 67 and a VL comprising the amino acid sequence of SEQ ID NO: 77. In one embodiment, the anti-PD-L1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 81 and a VL comprising the amino acid sequence of SEQ ID NO: 85.

In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 68, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 68. In one embodiment, the antibody molecule comprises a VL encoded by the nucleotide sequence of SEQ ID NO: 78, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 78. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 82, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 82. In one embodiment, the antibody molecule comprises a VL encoded by the nucleotide sequence of SEQ ID NO: 86, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 86. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 68 and a VL encoded by the nucleotide sequence of SEQ ID NO: 78. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 82 and a VL encoded by the nucleotide sequence of SEQ ID NO: 86.

In one embodiment, the anti-PD-L1 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 69, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 69. In one embodiment, the anti-PD-L1 antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 79, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 79. In one embodiment, the anti-PD-L1 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 83, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 83. In one embodiment, the anti-PD-L1 antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 87, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 87. In one embodiment, the anti-PD-L1 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 69 and a light chain comprising the amino acid sequence of SEQ ID NO: 79. In one embodiment, the anti-PD-L1 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 83 and a light chain comprising the amino acid sequence of SEQ ID NO: 87.

In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 76, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 76. In one embodiment, the antibody molecule comprises a light chain encoded by the nucleotide sequence of SEQ ID NO: 80, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 80. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 84, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 84. In one embodiment, the antibody molecule comprises a light chain encoded by the nucleotide sequence of SEQ ID NO: 88, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 88. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 76 and a light chain encoded by the nucleotide sequence of SEQ ID NO: 80. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 84 and a light chain encoded by the nucleotide sequence of SEQ ID NO: 88.

The antibody molecules described herein can be made by vectors, host cells, and methods described in US 2016/0108123, incorporated by reference in its entirety.

TABLE 5 Amino acid and nucleotide sequences of exemplary anti-PD-L1 antibody molecules BAP058-Clone O HC SEQ ID NO: 62 (Kabat) HCDR1 SYWMY SEQ ID NO: 63 (Kabat) HCDR2 RIDPNSGSTKYNEKFKN SEQ ID NO: 64 (Kabat) HCDR3 DYRKGLYAMDY SEQ ID NO: 65 HCDR1 GYTFTSY (Chothia) SEQ ID NO: 66 HCDR2 DPNSGS (Chothia) SEQ ID NO: 64 HCDR3 DYRKGLYAMDY (Chothia) SEQ ID NO: 67 VH EVQLVQSGAEVKKPGATVKISCKVSGYTFTSYWMYWVR QARGQRLEWIGRIDPNSGSTKYNEKFKNRFTISRDNSKNTL YLQMNSLRAEDTAVYYCARDYRKGLYAMDYWGQGTTV TVSS SEQ ID NO: 68 DNA GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAA VH ACCCGGCGCTACCGTGAAGATTAGCTGTAAAGTCTCAG GCTACACCTTCACTAGCTACTGGATGTACTGGGTCCGAC AGGCTAGAGGGCAAAGACTGGAGTGGATCGGTAGAATC GACCCTAATAGCGGCTCTACTAAGTATAACGAGAAGTT TAAGAATAGGTTCACTATTAGTAGGGATAACTCTAAGA ACACCCTGTACCTGCAGATGAATAGCCTGAGAGCCGAG GACACCGCCGTCTACTACTGCGCTAGAGACTATAGAAA GGGCCTGTACGCTATGGACTACTGGGGTCAAGGCACTA CCGTGACCGTGTCTTCA SEQ ID NO: 69 Heavy EVQLVQSGAEVKKPGATVKISCKVSGYTFTSYWMYWVR chain QARGQRLEWIGRIDPNSGSTKYNEKFKNRFTISRDNSKNTL YLQMNSLRAEDTAVYYCARDYRKGLYAMDYWGQGTTV TVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT KTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWY VDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHY TQKSLSLSLG SEQ ID NO: 76 DNA GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAA heavy ACCCGGCGCTACCGTGAAGATTAGCTGTAAAGTCTCAG chain GCTACACCTTCACTAGCTACTGGATGTACTGGGTCCGAC AGGCTAGAGGGCAAAGACTGGAGTGGATCGGTAGAATC GACCCTAATAGCGGCTCTACTAAGTATAACGAGAAGTT TAAGAATAGGTTCACTATTAGTAGGGATAACTCTAAGA ACACCCTGTACCTGCAGATGAATAGCCTGAGAGCCGAG GACACCGCCGTCTACTACTGCGCTAGAGACTATAGAAA GGGCCTGTACGCTATGGACTACTGGGGTCAAGGCACTA CCGTGACCGTGTCTTCAGCTAGCACTAAGGGCCCGTCCG TGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAAT CCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCC CGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTG ACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGC TCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCT TCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTG GACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGT CGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCG CGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCC CACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACC CCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGA AGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCG TCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGA GCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGAC GGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACA AGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATC GAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGG AACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAA ATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAG GGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTC CAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTC CGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGC GGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAA TGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAA CCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA BAP058-Clone O LC SEQ ID NO: 70 (Kabat) LCDR1 KASQDVGTAVA SEQ ID NO: 71 (Kabat) LCDR2 WASTRHT SEQ ID NO: 72 (Kabat) LCDR3 QQYNSYPLT SEQ ID NO: 73 LCDR1 SQDVGTA (Chothia) SEQ ID NO: 74 LCDR2 WAS (Chothia) SEQ ID NO: 75 LCDR3 YNSYPL (Chothia) SEQ ID NO: 77 VL AIQLTQSPSSLSASVGDRVTITCKASQDVGTAVAWYLQKP GQSPQLLIYWASTRHTGVPSRFSGSGSGTDFTFTISSLEAED AATYYCQQYNSYPLTFGQGTKVEIK SEQ ID NO: 78 DNA GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCT VL AGTGTGGGCGATAGAGTGACTATCACCTGTAAAGCCTC TCAGGACGTGGGCACCGCCGTGGCCTGGTATCTGCAGA AGCCTGGTCAATCACCTCAGCTGCTGATCTACTGGGCCT CTACTAGACACACCGGCGTGCCCTCTAGGTTTAGCGGTA GCGGTAGTGGCACCGACTTCACCTTCACTATCTCTTCAC TGGAAGCCGAGGACGCCGCTACCTACTACTGTCAGCAG TATAATAGCTACCCCCTGACCTTCGGTCAAGGCACTAAG GTCGAGATTAAG SEQ ID NO: 79 Light AIQLTQSPSSLSASVGDRVTITCKASQDVGTAVAWYLQKP chain GQSPQLLIYWASTRHTGVPSRFSGSGSGTDFTFTISSLEAED AATYYCQQYNSYPLTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS SPVTKSFNRGEC SEQ ID NO: 80 DNA GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCT light AGTGTGGGCGATAGAGTGACTATCACCTGTAAAGCCTC chain TCAGGACGTGGGCACCGCCGTGGCCTGGTATCTGCAGA AGCCTGGTCAATCACCTCAGCTGCTGATCTACTGGGCCT CTACTAGACACACCGGCGTGCCCTCTAGGTTTAGCGGTA GCGGTAGTGGCACCGACTTCACCTTCACTATCTCTTCAC TGGAAGCCGAGGACGCCGCTACCTACTACTGTCAGCAG TATAATAGCTACCCCCTGACCTTCGGTCAAGGCACTAAG GTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTT CATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCA CCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCC GGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCT GCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAG GACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCT GACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGT ACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCC GTGACCAAGAGCTTCAACAGGGGCGAGTGC BAP058-Clone N HC SEQ ID NO: 62 (Kabat) HCDR1 SYWMY SEQ ID NO: 63 (Kabat) HCDR2 RIDPNSGSTKYNEKFKN SEQ ID NO: 64 (Kabat) HCDR3 DYRKGLYAMDY SEQ ID NO: 65 HCDR1 GYTFTSY (Chothia) SEQ ID NO: 66 HCDR2 DPNSGS (Chothia) SEQ ID NO: 64 HCDR3 DYRKGLYAMDY (Chothia) SEQ ID NO: 81 VH EVQLVQSGAEVKKPGATVKISCKVSGYTFTSYWMYWVR QATGQGLEWMGRIDPNSGSTKYNEKFKNRVTITADKSTST AYMELSSLRSEDTAVYYCARDYRKGLYAMDYWGQGTTV TVSS SEQ ID NO: 82 DNA GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAA VH ACCCGGCGCTACCGTGAAGATTAGCTGTAAAGTCTCAG GCTACACCTTCACTAGCTACTGGATGTACTGGGTCCGAC AGGCTACCGGTCAAGGCCTGGAGTGGATGGGTAGAATC GACCCTAATAGCGGCTCTACTAAGTATAACGAGAAGTT TAAGAATAGAGTGACTATCACCGCCGATAAGTCTACTA GCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAG GACACCGCCGTCTACTACTGCGCTAGAGACTATAGAAA GGGCCTGTACGCTATGGACTACTGGGGTCAAGGCACTA CCGTGACCGTGTCTTCA SEQ ID NO: 83 Heavy EVQLVQSGAEVKKPGATVKISCKVSGYTFTSYWMYWVR chain QATGQGLEWMGRIDPNSGSTKYNEKFKNRVTITADKSTST AYMELSSLRSEDTAVYYCARDYRKGLYAMDYWGQGTTV TVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT KTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWY VDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHY TQKSLSLSLG SEQ ID NO: 84 DNA GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAA heavy ACCCGGCGCTACCGTGAAGATTAGCTGTAAAGTCTCAG chain GCTACACCTTCACTAGCTACTGGATGTACTGGGTCCGAC AGGCTACCGGTCAAGGCCTGGAGTGGATGGGTAGAATC GACCCTAATAGCGGCTCTACTAAGTATAACGAGAAGTT TAAGAATAGAGTGACTATCACCGCCGATAAGTCTACTA GCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAG GACACCGCCGTCTACTACTGCGCTAGAGACTATAGAAA GGGCCTGTACGCTATGGACTACTGGGGTCAAGGCACTA CCGTGACCGTGTCTTCAGCTAGCACTAAGGGCCCGTCCG TGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAAT CCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCC CGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTG ACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGC TCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCT TCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTG GACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGT CGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCG CGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCC CACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACC CCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGA AGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCG TCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGA GCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGAC GGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACA AGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATC GAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGG AACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAA ATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAG GGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTC CAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTC CGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGC GGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAA TGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAA CCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA BAP058-Clone N LC SEQ ID NO: 70 (Kabat) LCDR1 KASQDVGTAVA SEQ ID NO: 71 (Kabat) LCDR2 WASTRHT SEQ ID NO: 72 (Kabat) LCDR3 QQYNSYPLT SEQ ID NO: 73 LCDR1 SQDVGTA (Chothia) SEQ ID NO: 74 LCDR2 WAS (Chothia) SEQ ID NO: 75 LCDR3 YNSYPL (Chothia) SEQ ID NO: 85 VL DVVMTQSPLSLPVTLGQPASISCKASQDVGTAVAWYQQK PGQAPRLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPD DFATYYCQQYNSYPLTFGQGTKVEIK SEQ ID NO: 86 DNA GACGTCGTGATGACTCAGTCACCCCTGAGCCTGCCCGTG VL ACCCTGGGGCAGCCCGCCTCTATTAGCTGTAAAGCCTCT CAGGACGTGGGCACCGCCGTGGCCTGGTATCAGCAGAA GCCAGGGCAAGCCCCTAGACTGCTGATCTACTGGGCCT CTACTAGACACACCGGCGTGCCCTCTAGGTTTAGCGGTA GCGGTAGTGGCACCGAGTTCACCCTGACTATCTCTTCAC TGCAGCCCGACGACTTCGCTACCTACTACTGTCAGCAGT ATAATAGCTACCCCCTGACCTTCGGTCAAGGCACTAAG GTCGAGATTAAG SEQ ID NO: 87 Light DVVMTQSPLSLPVTLGQPASISCKASQDVGTAVAWYQQK chain PGQAPRLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPD DFATYYCQQYNSYPLTFGQGTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC SEQ ID NO: 88 DNA GACGTCGTGATGACTCAGTCACCCCTGAGCCTGCCCGTG light ACCCTGGGGCAGCCCGCCTCTATTAGCTGTAAAGCCTCT chain CAGGACGTGGGCACCGCCGTGGCCTGGTATCAGCAGAA GCCAGGGCAAGCCCCTAGACTGCTGATCTACTGGGCCT CTACTAGACACACCGGCGTGCCCTCTAGGTTTAGCGGTA GCGGTAGTGGCACCGAGTTCACCCTGACTATCTCTTCAC TGCAGCCCGACGACTTCGCTACCTACTACTGTCAGCAGT ATAATAGCTACCCCCTGACCTTCGGTCAAGGCACTAAG GTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTT CATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCA CCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCC GGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCT GCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAG GACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCT GACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGT ACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCC GTGACCAAGAGCTTCAACAGGGGCGAGTGC BAP058-Clone O HC SEQ ID NO: 89 (Kabat) HCDR1 agctactggatgtac SEQ ID NO: 90 (Kabat) HCDR2 agaatcgaccctaatagcggctctactaagtataacgagaagtttaagaat SEQ ID NO: 91 (Kabat) HCDR3 gactatagaaagggcctgtacgctatggactac SEQ ID NO: 92 HCDR1 ggctacaccttcactagctac (Chothia) SEQ ID NO: 93 HCDR2 gaccctaatagcggctct (Chothia) SEQ ID NO: 91 HCDR3 gactatagaaagggcctgtacgctatggactac (Chothia) BAP058-Clone O LC SEQ ID NO: 94 (Kabat) LCDR1 aaagcctctcaggacgtgggcaccgccgtggcc SEQ ID NO: 95 (Kabat) LCDR2 tgggcctctactagacacacc SEQ ID NO: 96 (Kabat) LCDR3 cagcagtataatagctaccccctgacc SEQ ID NO: 97 LCDR1 tctcaggacgtgggcaccgcc (Chothia) SEQ ID NO: 98 LCDR2 tgggcctct (Chothia) SEQ ID NO: 99 LCDR3 tataatagctaccccctg (Chothia) BAP058-Clone N HC SEQ ID NO: 89 (Kabat) HCDR1 agctactggatgtac SEQ ID NO: 90 (Kabat) HCDR2 agaatcgaccctaatagcggctctactaagtataacgagaagtttaagaat SEQ ID NO: 91 (Kabat) HCDR3 gactatagaaagggcctgtacgctatggactac SEQ ID NO: 92 HCDR1 ggctacaccttcactagctac (Chothia) SEQ ID NO: 93 HCDR2 gaccctaatagcggctct (Chothia) SEQ ID NO: 91 HCDR3 gactatagaaagggcctgtacgctatggactac (Chothia) BAP058-Clone N LC SEQ ID NO: 94 (Kabat) LCDR1 aaagcctctcaggacgtgggcaccgccgtggcc SEQ ID NO: 95 (Kabat) LCDR2 tgggcctctactagacacacc SEQ ID NO: 96 (Kabat) LCDR3 cagcagtataatagctaccccctgacc SEQ ID NO: 97 LCDR1 tctcaggacgtgggcaccgcc (Chothia) SEQ ID NO: 98 LCDR2 tgggcctct (Chothia) SEQ ID NO: 99 LCDR3 tataatagctaccccctg (Chothia) Other Exemplary PD-L1 Inhibitors

In some embodiments, the PD-L1 inhibitor is anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 inhibitor is selected from YW243.55.S70, MPDL3280A, MEDI-4736, or MDX-1105MSB-0010718C (also referred to as A09-246-2) disclosed in, e.g., WO 2013/0179174, and having a sequence disclosed herein (or a sequence substantially identical or similar thereto, e.g., a sequence at least 85%, 90%, 95% identical or higher to the sequence specified).

In one embodiment, the PD-L1 inhibitor is MDX-1105. MDX-1105, also known as BMS-936559, is an anti-PD-L1 antibody described in PCT Publication No. WO 2007/005874.

In one embodiment, the PD-L1 inhibitor is YW243.55.S70. The YW243.55.S70 antibody is an anti-PD-L1 described in PCT Publication No. WO 2010/077634.

In one embodiment, the PD-L1 inhibitor is MDPL3280A (Genentech/Roche) also known as Atezolizumabm, RG7446, RO5541267, YW243.55.S70, or TECENTRIQ™. MDPL3280A is a human Fc optimized IgG1 monoclonal antibody that binds to PD-L1. MDPL3280A and other human monoclonal antibodies to PD-L1 are disclosed in U.S. Pat. No. 7,943,743 and U.S Publication No.: 20120039906 incorporated by reference in its entirety. In one embodiment, the anti-PD-L1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of Atezolizumab, e.g., as disclosed in Table 6.

In other embodiments, the PD-L2 inhibitor is AMP-224. AMP-224 is a PD-L2 Fc fusion soluble receptor that blocks the interaction between PD1 and B7-H1 (B7-DCIg; Amplimmune; e.g., disclosed in PCT Publication Nos. WO2010/027827 and WO2011/066342).

In one embodiment, the PD-L1 inhibitor is an anti-PD-L1 antibody molecule. In one embodiment, the anti-PD-L1 antibody molecule is Avelumab (Merck Serono and Pfizer), also known as MSB0010718C. Avelumab and other anti-PD-L1 antibodies are disclosed in WO 2013/079174, incorporated by reference in its entirety. In one embodiment, the anti-PD-L1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of Avelumab, e.g., as disclosed in Table 6.

In one embodiment, the anti-PD-L1 antibody molecule is Durvalumab (MedImmune/AstraZeneca), also known as MEDI4736. Durvalumab and other anti-PD-L1 antibodies are disclosed in U.S. Pat. No. 8,779,108, incorporated by reference in its entirety. In one embodiment, the anti-PD-L1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of Durvalumab, e.g., as disclosed in Table 6.

In one embodiment, the anti-PD-L1 antibody molecule is BMS-936559 (Bristol-Myers Squibb), also known as MDX-1105 or 12A4. BMS-936559 and other anti-PD-L1 antibodies are disclosed in U.S. Pat. No. 7,943,743 and WO 2015/081158, incorporated by reference in their entirety. In one embodiment, the anti-PD-L1 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of BMS-936559, e.g., as disclosed in Table 6.

Further known anti-PD-L1 antibodies include those described, e.g., in WO 2015/181342, WO 2014/100079, WO 2016/000619, WO 2014/022758, WO 2014/055897, WO 2015/061668, WO 2013/079174, WO 2012/145493, WO 2015/112805, WO 2015/109124, WO 2015/195163, U.S. Pat. Nos. 8,168,179, 8,552,154, 8,460,927, and 9,175,082, incorporated by reference in their entirety.

In one embodiment, the anti-PD-L1 antibody is an antibody that competes for binding with, and/or binds to the same epitope on PD-L1 as, one of the anti-PD-L1 antibodies described herein.

TABLE 6 Amino acid sequences of other exemplary anti-PD-L1 antibody molecules Atezolizumab SEQ ID NO: Heavy EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLE 100 chain WVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTA VYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCK VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: Light DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKL 101 chain LIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHP ATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGEC Avelumab SEQ ID NO: Heavy EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLE 102 chain WVSSIYPSGGITFYADTVKGRFTISRDNSKNTLYLQMNSLRAEDTAV YYCARIKLGTVTTVDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK VSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: Light QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPK 103 chain LMIYDVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTS SSTRVFGTGTKVTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDF YPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQ WKSHRSYSCQVTHEGSTVEKTVAPTECS Durvalumab SEQ ID NO: Heavy EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGL 104 chain EWVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDT AVYYCAREGGWFGELAFDYWGQGTLVTVSSASTKGPSVFPLAPSSK STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPASIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: Light EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLL 105 chain IYDASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLP WTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGEC BMS-936559 SEQ ID NO: VH QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQGLE 106 WMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRSEDTAV YFCARKFHFVSGSPFGMDVWGQGTTVTVSS SEQ ID NO: VL EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLI 107 YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPT FGQGTKVEIK LAG-3 Inhibitors

In some embodiments, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with a LAG-3 inhibitor to treat a disease, e.g., cancer. In some embodiments, the LAG-3 inhibitor is selected from LAG525 (Novartis), BMS-986016 (Bristol-Myers Squibb), or TSR-033 (Tesaro).

Exemplary LAG-3 Inhibitors

In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody molecule. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody molecule as disclosed in US 2015/0259420, published on Sep. 17, 2015, entitled “Antibody Molecules to LAG-3 and Uses Thereof,” incorporated by reference in its entirety.

In one embodiment, the anti-LAG-3 antibody molecule comprises at least one, two, three, four, five or six complementarity determining regions (CDRs) (or collectively all of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 7 (e.g., from the heavy and light chain variable region sequences of BAP050-Clone I or BAP050-Clone J disclosed in Table 7), or encoded by a nucleotide sequence shown in Table 7. In some embodiments, the CDRs are according to the Kabat definition (e.g., as set out in Table 7). In some embodiments, the CDRs are according to the Chothia definition (e.g., as set out in Table 7). In some embodiments, the CDRs are according to the combined CDR definitions of both Kabat and Chothia (e.g., as set out in Table 7). In one embodiment, the combination of Kabat and Chothia CDR of VH CDR1 comprises the amino acid sequence GFTLTNYGMN (SEQ ID NO: 173). In one embodiment, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions (e.g., conservative amino acid substitutions) or deletions, relative to an amino acid sequence shown in Table 7, or encoded by a nucleotide sequence shown in Table 7.

In one embodiment, the anti-LAG-3 antibody molecule comprises a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence of SEQ ID NO: 108, a VHCDR2 amino acid sequence of SEQ ID NO: 109, and a VHCDR3 amino acid sequence of SEQ ID NO: 110; and a light chain variable region (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 117, a VLCDR2 amino acid sequence of SEQ ID NO: 118, and a VLCDR3 amino acid sequence of SEQ ID NO: 119, each disclosed in Table 7.

In one embodiment, the anti-LAG-3 antibody molecule comprises a VH comprising a VHCDR1 encoded by the nucleotide sequence of SEQ ID NO: 143 or 144, a VHCDR2 encoded by the nucleotide sequence of SEQ ID NO: 145 or 146, and a VHCDR3 encoded by the nucleotide sequence of SEQ ID NO: 147 or 148; and a VL comprising a VLCDR1 encoded by the nucleotide sequence of SEQ ID NO: 153 or 154, a VLCDR2 encoded by the nucleotide sequence of SEQ ID NO: 155 or 156, and a VLCDR3 encoded by the nucleotide sequence of SEQ ID NO: 157 or 158, each disclosed in Table 7. In one embodiment, the anti-LAG-3 antibody molecule comprises a VH comprising a VHCDR1 encoded by the nucleotide sequence of SEQ ID NO: 165 or 144, a VHCDR2 encoded by the nucleotide sequence of SEQ ID NO: 166 or 146, and a VHCDR3 encoded by the nucleotide sequence of SEQ ID NO: 167 or 148; and a VL comprising a VLCDR1 encoded by the nucleotide sequence of SEQ ID NO: 153 or 154, a VLCDR2 encoded by the nucleotide sequence of SEQ ID NO: 155 or 156, and a VLCDR3 encoded by the nucleotide sequence of SEQ ID NO: 157 or 158, each disclosed in Table 7.

In one embodiment, the anti-LAG-3 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 113, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 113. In one embodiment, the anti-LAG-3 antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 125, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 125. In one embodiment, the anti-LAG-3 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 131, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 131. In one embodiment, the anti-LAG-3 antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 137, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 137. In one embodiment, the anti-LAG-3 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 113 and a VL comprising the amino acid sequence of SEQ ID NO: 125. In one embodiment, the anti-LAG-3 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 131 and a VL comprising the amino acid sequence of SEQ ID NO: 137.

In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 114 or 115, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 114 or 115. In one embodiment, the antibody molecule comprises a VL encoded by the nucleotide sequence of SEQ ID NO: 126 or 127, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 126 or 127. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 132 or 133, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 132 or 133. In one embodiment, the antibody molecule comprises a VL encoded by the nucleotide sequence of SEQ ID NO: 138 or 139, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 138 or 139. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 114 or 115 and a VL encoded by the nucleotide sequence of SEQ ID NO: 126 or 127. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 132 or 133 and a VL encoded by the nucleotide sequence of SEQ ID NO: 138 or 139.

In one embodiment, the anti-LAG-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 116, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 116. In one embodiment, the anti-LAG-3 antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 128, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 128. In one embodiment, the anti-LAG-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 134, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 134. In one embodiment, the anti-LAG-3 antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 140, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 140. In one embodiment, the anti-LAG-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 116 and a light chain comprising the amino acid sequence of SEQ ID NO: 128. In one embodiment, the anti-LAG-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 134 and a light chain comprising the amino acid sequence of SEQ ID NO: 140.

In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 123 or 124, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 123 or 124. In one embodiment, the antibody molecule comprises a light chain encoded by the nucleotide sequence of SEQ ID NO: 129 or 130, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 129 or 130. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 135 or 136, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 135 or 136. In one embodiment, the antibody molecule comprises a light chain encoded by the nucleotide sequence of SEQ ID NO: 141 or 142, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 141 or 142. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 123 or 124 and a light chain encoded by the nucleotide sequence of SEQ ID NO: 129 or 130. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 135 or 136 and a light chain encoded by the nucleotide sequence of SEQ ID NO: 141 or 142.

The antibody molecules described herein can be made by vectors, host cells, and methods described in US 2015/0259420, incorporated by reference in its entirety.

TABLE 7 Amino acid and nucleotide sequences of exemplary anti-LAG-3 antibody molecules BAP050-Clone I HC SEQ ID NO: 108 HCDR1 NYGMN (Kabat) SEQ ID NO: 109 HCDR2 WINTDTGEPTYADDFKG (Kabat) SEQ ID NO: 110 HCDR3 NPPYYYGTNNAEAMDY (Kabat) SEQ ID NO: 111 HCDR1 GFTLTNY (Chothia) SEQ ID NO: 112 HCDR2 NTDTGE (Chothia) SEQ ID NO: 110 HCDR3 NPPYYYGTNNAEAMDY (Chothia) SEQ ID NO: 113 VH QVQLVQSGAEVKKPGASVKVSCKASGFTLTNYGMNWVRQAR GQRLEWIGWINTDTGEPTYADDFKGRFVFSLDTSVSTAYLQISS LKAEDTAVYYCARNPPYYYGTNNAEAMDYWGQGTTVTVSS SEQ ID NO: 114 DNA CAAGTGCAGCTGGTGCAGTCGGGAGCCGAAGTGAAGAAGCC VH TGGAGCCTCGGTGAAGGTGTCGTGCAAGGCATCCGGATTCA CCCTCACCAATTACGGGATGAACTGGGTCAGACAGGCCCGG GGTCAACGGCTGGAGTGGATCGGATGGATTAACACCGACAC CGGGGAGCCTACCTACGCGGACGATTTCAAGGGACGGTTCG TGTTCTCCCTCGACACCTCCGTGTCCACCGCCTACCTCCAAA TCTCCTCACTGAAAGCGGAGGACACCGCCGTGTACTATTGC GCGAGGAACCCGCCCTACTACTACGGAACCAACAACGCCGA AGCCATGGACTACTGGGGCCAGGGCACCACTGTGACTGTGT CCAGC SEQ ID NO: 115 DNA CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACC VH TGGCGCCTCCGTGAAGGTGTCCTGCAAGGCCTCTGGCTTCAC CCTGACCAACTACGGCATGAACTGGGTGCGACAGGCCAGGG GCCAGCGGCTGGAATGGATCGGCTGGATCAACACCGACACC GGCGAGCCTACCTACGCCGACGACTTCAAGGGCAGATTCGT GTTCTCCCTGGACACCTCCGTGTCCACCGCCTACCTGCAGAT CTCCAGCCTGAAGGCCGAGGATACCGCCGTGTACTACTGCG CCCGGAACCCCCCTTACTACTACGGCACCAACAACGCCGAG GCCATGGACTATTGGGGCCAGGGCACCACCGTGACCGTGTC CTCT SEQ ID NO: 116 Heavy QVQLVQSGAEVKKPGASVKVSCKASGFTLTNYGMNWVRQAR chain GQRLEWIGWINTDTGEPTYADDFKGRFVFSLDTSVSTAYLQISS LKAEDTAVYYCARNPPYYYGTNNAEAMDYWGQGTTVTVSSA STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKP SNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKA KGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSC SVMHEALHNHYTQKSLSLSLG SEQ ID NO: 123 DNA CAAGTGCAGCTGGTGCAGTCGGGAGCCGAAGTGAAGAAGCC heavy TGGAGCCTCGGTGAAGGTGTCGTGCAAGGCATCCGGATTCA chain CCCTCACCAATTACGGGATGAACTGGGTCAGACAGGCCCGG GGTCAACGGCTGGAGTGGATCGGATGGATTAACACCGACAC CGGGGAGCCTACCTACGCGGACGATTTCAAGGGACGGTTCG TGTTCTCCCTCGACACCTCCGTGTCCACCGCCTACCTCCAAA TCTCCTCACTGAAAGCGGAGGACACCGCCGTGTACTATTGC GCGAGGAACCCGCCCTACTACTACGGAACCAACAACGCCGA AGCCATGGACTACTGGGGCCAGGGCACCACTGTGACTGTGT CCAGCGCGTCCACTAAGGGCCCGTCCGTGTTCCCCCTGGCAC CTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCT GCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCT GGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCC GCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTG GTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACT TGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAA GCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTC CCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCC CACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTG AAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCG GAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCA CAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCA CTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACT GGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAA GGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCA AGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCG AGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTG CCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATG GGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCC CTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGC GGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGT GTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTA CACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 124 DNA CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACC heavy TGGCGCCTCCGTGAAGGTGTCCTGCAAGGCCTCTGGCTTCAC chain CCTGACCAACTACGGCATGAACTGGGTGCGACAGGCCAGGG GCCAGCGGCTGGAATGGATCGGCTGGATCAACACCGACACC GGCGAGCCTACCTACGCCGACGACTTCAAGGGCAGATTCGT GTTCTCCCTGGACACCTCCGTGTCCACCGCCTACCTGCAGAT CTCCAGCCTGAAGGCCGAGGATACCGCCGTGTACTACTGCG CCCGGAACCCCCCTTACTACTACGGCACCAACAACGCCGAG GCCATGGACTATTGGGGCCAGGGCACCACCGTGACCGTGTC CTCTGCTTCTACCAAGGGGCCCAGCGTGTTCCCCCTGGCCCC CTGCTCCAGAAGCACCAGCGAGAGCACAGCCGCCCTGGGCT GCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCT GGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCC GCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGT GGTGACCGTGCCCAGCAGCAGCCTGGGCACCAAGACCTACA CCTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGAC AAGAGGGTGGAGAGCAAGTACGGCCCACCCTGCCCCCCCTG CCCAGCCCCCGAGTTCCTGGGCGGACCCAGCGTGTTCCTGTT CCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGAACCC CCGAGGTGACCTGTGTGGTGGTGGACGTGTCCCAGGAGGAC CCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGT GCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTTTAAC AGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCA GGACTGGCTGAACGGCAAAGAGTACAAGTGTAAGGTCTCCA ACAAGGGCCTGCCAAGCAGCATCGAAAAGACCATCAGCAA GGCCAAGGGCCAGCCTAGAGAGCCCCAGGTCTACACCCTGC CACCCAGCCAAGAGGAGATGACCAAGAACCAGGTGTCCCTG ACCTGTCTGGTGAAGGGCTTCTACCCAAGCGACATCGCCGT GGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAG ACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTG TACAGCAGGCTGACCGTGGACAAGTCCAGATGGCAGGAGGG CAACGTCTTTAGCTGCTCCGTGATGCACGAGGCCCTGCACAA CCACTACACCCAGAAGAGCCTGAGCCTGTCCCTGGGC BAP050-Clone I LC SEQ ID NO: 117 LCDR1 SSSQDISNYLN (Kabat) SEQ ID NO: 118 LCDR2 YTSTLHL (Kabat) SEQ ID NO: 119 LCDR3 QQYYNLPWT (Kabat) SEQ ID NO: 120 LCDR1 SQDISNY (Chothia) SEQ ID NO: 121 LCDR2 YTS (Chothia) SEQ ID NO: 122 LCDR3 YYNLPW (Chothia) SEQ ID NO: 125 VL DIQMTQSPSSLSASVGDRVTITCSSSQDISNYLNWYLQKPGQSP QLLIYYTSTLHLGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQ QYYNLPWTFGQGTKVEIK SEQ ID NO: 126 DNA GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGT VL GTGGGCGATAGAGTGACTATCACCTGTAGCTCTAGTCAGGA TATCTCTAACTACCTGAACTGGTATCTGCAGAAGCCCGGTCA ATCACCTCAGCTGCTGATCTACTACACTAGCACCCTGCACCT GGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCG AGTTCACCCTGACTATCTCTAGCCTGCAGCCCGACGACTTCG CTACCTACTACTGTCAGCAGTACTATAACCTGCCCTGGACCT TCGGTCAAGGCACTAAGGTCGAGATTAAG SEQ ID NO: 127 DNA GACATCCAGATGACCCAGTCCCCCTCCAGCCTGTCTGCTTCC VL GTGGGCGACAGAGTGACCATCACCTGTTCCTCCAGCCAGGA CATCTCCAACTACCTGAACTGGTATCTGCAGAAGCCCGGCC AGTCCCCTCAGCTGCTGATCTACTACACCTCCACCCTGCACC TGGGCGTGCCCTCCAGATTTTCCGGCTCTGGCTCTGGCACCG AGTTTACCCTGACCATCAGCTCCCTGCAGCCCGACGACTTCG CCACCTACTACTGCCAGCAGTACTACAACCTGCCCTGGACCT TCGGCCAGGGCACCAAGGTGGAAATCAAG SEQ ID NO: 128 Light DIQMTQSPSSLSASVGDRVTITCSSSQDISNYLNWYLQKPGQSP chain QLLIYYTSTLHLGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQ QYYNLPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 129 DNA GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGT light GTGGGCGATAGAGTGACTATCACCTGTAGCTCTAGTCAGGA chain TATCTCTAACTACCTGAACTGGTATCTGCAGAAGCCCGGTCA ATCACCTCAGCTGCTGATCTACTACACTAGCACCCTGCACCT GGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCG AGTTCACCCTGACTATCTCTAGCCTGCAGCCCGACGACTTCG CTACCTACTACTGTCAGCAGTACTATAACCTGCCCTGGACCT TCGGTCAAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCC GCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTG AAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTT CTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACG CCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCA GGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGA CCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCC TGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAA GAGCTTCAACAGGGGCGAGTGC SEQ ID NO: 130 DNA GACATCCAGATGACCCAGTCCCCCTCCAGCCTGTCTGCTTCC light GTGGGCGACAGAGTGACCATCACCTGTTCCTCCAGCCAGGA chain CATCTCCAACTACCTGAACTGGTATCTGCAGAAGCCCGGCC AGTCCCCTCAGCTGCTGATCTACTACACCTCCACCCTGCACC TGGGCGTGCCCTCCAGATTTTCCGGCTCTGGCTCTGGCACCG AGTTTACCCTGACCATCAGCTCCCTGCAGCCCGACGACTTCG CCACCTACTACTGCCAGCAGTACTACAACCTGCCCTGGACCT TCGGCCAGGGCACCAAGGTGGAAATCAAGCGTACGGTGGCC GCTCCCAGCGTGTTCATCTTCCCCCCAAGCGACGAGCAGCTG AAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTT CTACCCCAGGGAGGCCAAGGTGCAGTGGAAGGTGGACAAC GCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGC AGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTG ACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGC CTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCA AGAGCTTCAACAGGGGCGAGTGC BAP050-Clone J HC SEQ ID NO: 108 HCDR1 NYGMN (Kabat) SEQ ID NO: 109 HCDR2 WINTDTGEPTYADDFKG (Kabat) SEQ ID NO: 110 HCDR3 NPPYYYGTNNAEAMDY (Kabat) SEQ ID NO: 111 HCDR1 GFTLTNY (Chothia) SEQ ID NO: 112 HCDR2 NTDTGE (Chothia) SEQ ID NO: 110 HCDR3 NPPYYYGTNNAEAMDY (Chothia) SEQ ID NO: 131 VH QVQLVQSGAEVKKPGASVKVSCKASGFTLTNYGMNWVRQAP GQGLEWMGWINTDTGEPTYADDFKGRFVFSLDTSVSTAYLQIS SLKAEDTAVYYCARNPPYYYGTNNAEAMDYWGQGTTVTVSS SEQ ID NO: 132 DNA CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACC VH CGGCGCTAGTGTGAAAGTCAGCTGTAAAGCTAGTGGCTTCA CCCTGACTAACTACGGGATGAACTGGGTCCGCCAGGCCCCA GGTCAAGGCCTCGAGTGGATGGGCTGGATTAACACCGACAC CGGCGAGCCTACCTACGCCGACGACTTTAAGGGCAGATTCG TGTTTAGCCTGGACACTAGTGTGTCTACCGCCTACCTGCAGA TCTCTAGCCTGAAGGCCGAGGACACCGCCGTCTACTACTGC GCTAGAAACCCCCCCTACTACTACGGCACTAACAACGCCGA GGCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGT CTAGC SEQ ID NO: 133 DNA CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACC VH TGGCGCCTCCGTGAAGGTGTCCTGCAAGGCCTCTGGCTTCAC CCTGACCAACTACGGCATGAACTGGGTGCGACAGGCCCCTG GACAGGGCCTGGAATGGATGGGCTGGATCAACACCGACACC GGCGAGCCTACCTACGCCGACGACTTCAAGGGCAGATTCGT GTTCTCCCTGGACACCTCCGTGTCCACCGCCTACCTGCAGAT CTCCAGCCTGAAGGCCGAGGATACCGCCGTGTACTACTGCG CCCGGAACCCCCCTTACTACTACGGCACCAACAACGCCGAG GCCATGGACTATTGGGGCCAGGGCACCACCGTGACCGTGTC CTCT SEQ ID NO: 134 Heavy QVQLVQSGAEVKKPGASVKVSCKASGFTLTNYGMNWVRQAP chain GQGLEWMGWINTDTGEPTYADDFKGRFVFSLDTSVSTAYLQIS SLKAEDTAVYYCARNPPYYYGTNNAEAMDYWGQGTTVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHK PSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQ FNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISK AKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFS CSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 135 DNA CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACC heavy CGGCGCTAGTGTGAAAGTCAGCTGTAAAGCTAGTGGCTTCA chain CCCTGACTAACTACGGGATGAACTGGGTCCGCCAGGCCCCA GGTCAAGGCCTCGAGTGGATGGGCTGGATTAACACCGACAC CGGCGAGCCTACCTACGCCGACGACTTTAAGGGCAGATTCG TGTTTAGCCTGGACACTAGTGTGTCTACCGCCTACCTGCAGA TCTCTAGCCTGAAGGCCGAGGACACCGCCGTCTACTACTGC GCTAGAAACCCCCCCTACTACTACGGCACTAACAACGCCGA GGCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGT CTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCAC CTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCT GCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCT GGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCC GCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTG GTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACT TGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAA GCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTC CCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCC CACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTG AAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCG GAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCA CAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCA CTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACT GGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAA GGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCA AGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCG AGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTG CCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATG GGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCC CTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGC GGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGT GTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTA CACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 136 DNA CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACC heavy TGGCGCCTCCGTGAAGGTGTCCTGCAAGGCCTCTGGCTTCAC chain CCTGACCAACTACGGCATGAACTGGGTGCGACAGGCCCCTG GACAGGGCCTGGAATGGATGGGCTGGATCAACACCGACACC GGCGAGCCTACCTACGCCGACGACTTCAAGGGCAGATTCGT GTTCTCCCTGGACACCTCCGTGTCCACCGCCTACCTGCAGAT CTCCAGCCTGAAGGCCGAGGATACCGCCGTGTACTACTGCG CCCGGAACCCCCCTTACTACTACGGCACCAACAACGCCGAG GCCATGGACTATTGGGGCCAGGGCACCACCGTGACCGTGTC CTCTGCTTCTACCAAGGGGCCCAGCGTGTTCCCCCTGGCCCC CTGCTCCAGAAGCACCAGCGAGAGCACAGCCGCCCTGGGCT GCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCT GGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCC GCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGT GGTGACCGTGCCCAGCAGCAGCCTGGGCACCAAGACCTACA CCTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGAC AAGAGGGTGGAGAGCAAGTACGGCCCACCCTGCCCCCCCTG CCCAGCCCCCGAGTTCCTGGGCGGACCCAGCGTGTTCCTGTT CCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGAACCC CCGAGGTGACCTGTGTGGTGGTGGACGTGTCCCAGGAGGAC CCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGT GCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTTTAAC AGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCA GGACTGGCTGAACGGCAAAGAGTACAAGTGTAAGGTCTCCA ACAAGGGCCTGCCAAGCAGCATCGAAAAGACCATCAGCAA GGCCAAGGGCCAGCCTAGAGAGCCCCAGGTCTACACCCTGC CACCCAGCCAAGAGGAGATGACCAAGAACCAGGTGTCCCTG ACCTGTCTGGTGAAGGGCTTCTACCCAAGCGACATCGCCGT GGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAG ACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTG TACAGCAGGCTGACCGTGGACAAGTCCAGATGGCAGGAGGG CAACGTCTTTAGCTGCTCCGTGATGCACGAGGCCCTGCACAA CCACTACACCCAGAAGAGCCTGAGCCTGTCCCTGGGC BAP050-Clone J LC SEQ ID NO: 117 LCDR1 SSSQDISNYLN (Kabat) SEQ ID NO: 118 LCDR2 YTSTLHL (Kabat) SEQ ID NO: 119 LCDR3 QQYYNLPWT (Kabat) SEQ ID NO: 120 LCDR1 SQDISNY (Chothia) SEQ ID NO: 121 LCDR2 YTS (Chothia) SEQ ID NO: 122 LCDR3 YYNLPW (Chothia) SEQ ID NO: 137 VL DIQMTQSPSSLSASVGDRVTITCSSSQDISNYLNWYQQKPGKAP KLLIYYTSTLHLGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQ QYYNLPWTFGQGTKVEIK SEQ ID NO: 138 DNA GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGT VL GTGGGCGATAGAGTGACTATCACCTGTAGCTCTAGTCAGGA TATCTCTAACTACCTGAACTGGTATCAGCAGAAGCCCGGTA AAGCCCCTAAGCTGCTGATCTACTACACTAGCACCCTGCACC TGGGAATCCCCCCTAGGTTTAGCGGTAGCGGCTACGGCACC GACTTCACCCTGACTATTAACAATATCGAGTCAGAGGACGC CGCCTACTACTTCTGTCAGCAGTACTATAACCTGCCCTGGAC CTTCGGTCAAGGCACTAAGGTCGAGATTAAG SEQ ID NO: 139 DNA GACATCCAGATGACCCAGTCCCCCTCCAGCCTGTCTGCTTCC VL GTGGGCGACAGAGTGACCATCACCTGTTCCTCCAGCCAGGA CATCTCCAACTACCTGAACTGGTATCAGCAGAAGCCCGGCA AGGCCCCCAAGCTGCTGATCTACTACACCTCCACCCTGCACC TGGGCATCCCCCCTAGATTCTCCGGCTCTGGCTACGGCACCG ACTTCACCCTGACCATCAACAACATCGAGTCCGAGGACGCC GCCTACTACTTCTGCCAGCAGTACTACAACCTGCCCTGGACC TTCGGCCAGGGCACCAAGGTGGAAATCAAG SEQ ID NO: 140 Light DIQMTQSPSSLSASVGDRVTITCSSSQDISNYLNWYQQKPGKAP chain KLLIYYTSTLHLGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQ QYYNLPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 141 DNA GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGT light GTGGGCGATAGAGTGACTATCACCTGTAGCTCTAGTCAGGA chain TATCTCTAACTACCTGAACTGGTATCAGCAGAAGCCCGGTA AAGCCCCTAAGCTGCTGATCTACTACACTAGCACCCTGCACC TGGGAATCCCCCCTAGGTTTAGCGGTAGCGGCTACGGCACC GACTTCACCCTGACTATTAACAATATCGAGTCAGAGGACGC CGCCTACTACTTCTGTCAGCAGTACTATAACCTGCCCTGGAC CTTCGGTCAAGGCACTAAGGTCGAGATTAAGCGTACGGTGG CCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGC TGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAAC TTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAA CGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGC AGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTG ACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGC CTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCA AGAGCTTCAACAGGGGCGAGTGC SEQ ID NO: 142 DNA GACATCCAGATGACCCAGTCCCCCTCCAGCCTGTCTGCTTCC light GTGGGCGACAGAGTGACCATCACCTGTTCCTCCAGCCAGGA chain CATCTCCAACTACCTGAACTGGTATCAGCAGAAGCCCGGCA AGGCCCCCAAGCTGCTGATCTACTACACCTCCACCCTGCACC TGGGCATCCCCCCTAGATTCTCCGGCTCTGGCTACGGCACCG ACTTCACCCTGACCATCAACAACATCGAGTCCGAGGACGCC GCCTACTACTTCTGCCAGCAGTACTACAACCTGCCCTGGACC TTCGGCCAGGGCACCAAGGTGGAAATCAAGCGTACGGTGGC CGCTCCCAGCGTGTTCATCTTCCCCCCAAGCGACGAGCAGCT GAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACT TCTACCCCAGGGAGGCCAAGGTGCAGTGGAAGGTGGACAAC GCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGC AGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTG ACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGC CTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCA AGAGCTTCAACAGGGGCGAGTGC BAP050-Clone I HC SEQ ID NO: 143 HCDR1 AATTACGGGATGAAC (Kabat) SEQ ID NO: 144 HCDR1 AACTACGGCATGAAC (Kabat) SEQ ID NO: 145 HCDR2 TGGATTAACACCGACACCGGGGAGCCTACCTACGCGGACGA (Kabat) TTTCAAGGGA SEQ ID NO: 146 HCDR2 TGGATCAACACCGACACCGGCGAGCCTACCTACGCCGACGA (Kabat) CTTCAAGGGC SEQ ID NO: 147 HCDR3 AACCCGCCCTACTACTACGGAACCAACAACGCCGAAGCCAT (Kabat) GGACTAC SEQ ID NO: 148 HCDR3 AACCCCCCTTACTACTACGGCACCAACAACGCCGAGGCCAT (Kabat) GGACTAT SEQ ID NO: 149 HCDR1 GGATTCACCCTCACCAATTAC (Chothia) SEQ ID NO: 150 HCDR1 GGCTTCACCCTGACCAACTAC (Chothia) SEQ ID NO: 151 HCDR2 AACACCGACACCGGGGAG (Chothia) SEQ ID NO: 152 HCDR2 AACACCGACACCGGCGAG (Chothia) SEQ ID NO: 147 HCDR3 AACCCGCCCTACTACTACGGAACCAACAACGCCGAAGCCAT (Chothia) GGACTAC SEQ ID NO: 148 HCDR3 AACCCCCCTTACTACTACGGCACCAACAACGCCGAGGCCAT (Chothia) GGACTAT BAP050-Clone I LC SEQ ID NO: 153 LCDR1 AGCTCTAGTCAGGATATCTCTAACTACCTGAAC (Kabat) SEQ ID NO: 154 LCDR1 TCCTCCAGCCAGGACATCTCCAACTACCTGAAC (Kabat) SEQ ID NO: 155 LCDR2 TACACTAGCACCCTGCACCTG (Kabat) SEQ ID NO: 156 LCDR2 TACACCTCCACCCTGCACCTG (Kabat) SEQ ID NO: 157 LCDR3 CAGCAGTACTATAACCTGCCCTGGACC (Kabat) SEQ ID NO: 158 LCDR3 CAGCAGTACTACAACCTGCCCTGGACC (Kabat) SEQ ID NO: 159 LCDR1 AGTCAGGATATCTCTAACTAC (Chothia) SEQ ID NO: 160 LCDR1 AGCCAGGACATCTCCAACTAC (Chothia) SEQ ID NO: 161 LCDR2 TACACTAGC (Chothia) SEQ ID NO: 162 LCDR2 TACACCTCC (Chothia) SEQ ID NO: 163 LCDR3 TACTATAACCTGCCCTGG (Chothia) SEQ ID NO: 164 LCDR3 TACTACAACCTGCCCTGG (Chothia) BAP050-Clone J HC SEQ ID NO: 165 HCDR1 AACTACGGGATGAAC (Kabat) SEQ ID NO: 144 HCDR1 AACTACGGCATGAAC (Kabat) SEQ ID NO: 166 HCDR2 TGGATTAACACCGACACCGGCGAGCCTACCTACGCCGACGA (Kabat) CTTTAAGGGC SEQ ID NO: 146 HCDR2 TGGATCAACACCGACACCGGCGAGCCTACCTACGCCGACGA (Kabat) CTTCAAGGGC SEQ ID NO: 167 HCDR3 AACCCCCCCTACTACTACGGCACTAACAACGCCGAGGCTAT (Kabat) GGACTAC SEQ ID NO: 148 HCDR3 AACCCCCCTTACTACTACGGCACCAACAACGCCGAGGCCAT (Kabat) GGACTAT SEQ ID NO: 168 HCDR1 GGCTTCACCCTGACTAACTAC (Chothia) SEQ ID NO: 150 HCDR1 GGCTTCACCCTGACCAACTAC (Chothia) SEQ ID NO: 151 HCDR2 AACACCGACACCGGGGAG (Chothia) SEQ ID NO: 152 HCDR2 AACACCGACACCGGCGAG (Chothia) SEQ ID NO: 167 HCDR3 AACCCCCCCTACTACTACGGCACTAACAACGCCGAGGCTAT (Chothia) GGACTAC SEQ ID NO: 148 HCDR3 AACCCCCCTTACTACTACGGCACCAACAACGCCGAGGCCAT (Chothia) GGACTAT BAP050-Clone J LC SEQ ID NO: 153 LCDR1 AGCTCTAGTCAGGATATCTCTAACTACCTGAAC (Kabat) SEQ ID NO: 154 LCDR1 TCCTCCAGCCAGGACATCTCCAACTACCTGAAC (Kabat) SEQ ID NO: 155 LCDR2 TACACTAGCACCCTGCACCTG (Kabat) SEQ ID NO: 156 LCDR2 TACACCTCCACCCTGCACCTG (Kabat) SEQ ID NO: 157 LCDR3 CAGCAGTACTATAACCTGCCCTGGACC (Kabat) SEQ ID NO: 158 LCDR3 CAGCAGTACTACAACCTGCCCTGGACC (Kabat) SEQ ID NO: 159 LCDR1 AGTCAGGATATCTCTAACTAC (Chothia) SEQ ID NO: 160 LCDR1 AGCCAGGACATCTCCAACTAC (Chothia) SEQ ID NO: 161 LCDR2 TACACTAGC (Chothia) SEQ ID NO: 162 LCDR2 TACACCTCC (Chothia) SEQ ID NO: 163 LCDR3 TACTATAACCTGCCCTGG (Chothia) SEQ ID NO: 164 LCDR3 TACTACAACCTGCCCTGG (Chothia) Other Exemplary LAG-3 Inhibitors

In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody molecule. In one embodiment, the LAG-3 inhibitor is BMS-986016 (Bristol-Myers Squibb), also known as BMS986016. BMS-986016 and other anti-LAG-3 antibodies are disclosed in WO 2015/116539 and U.S. Pat. No. 9,505,839, incorporated by reference in their entirety. In one embodiment, the anti-LAG-3 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of BMS-986016, e.g., as disclosed in Table 8.

In one embodiment, the anti-LAG-3 antibody molecule is TSR-033 (Tesaro). In one embodiment, the anti-LAG-3 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of TSR-033.

In one embodiment, the anti-LAG-3 antibody molecule is IMP731 or GSK2831781 (GSK and Prima BioMed). IMP731 and other anti-LAG-3 antibodies are disclosed in WO 2008/132601 and U.S. Pat. No. 9,244,059, incorporated by reference in their entirety. In one embodiment, the anti-LAG-3 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of IMP731, e.g., as disclosed in Table 8. In one embodiment, the anti-LAG-3 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of GSK2831781.

In one embodiment, the anti-LAG-3 antibody molecule is IMP761 (Prima BioMed). In one embodiment, the anti-LAG-3 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of IMP761.

Further known anti-LAG-3 antibodies include those described, e.g., in WO 2008/132601, WO 2010/019570, WO 2014/140180, WO 2015/116539, WO 2015/200119, WO 2016/028672, U.S. Pat. Nos. 9,244,059, 9,505,839, incorporated by reference in their entirety.

In one embodiment, the anti-LAG-3 antibody is an antibody that competes for binding with, and/or binds to the same epitope on LAG-3 as, one of the anti-LAG-3 antibodies described herein.

In one embodiment, the anti-LAG-3 inhibitor is a soluble LAG-3 protein, e.g., IMP321 (Prima BioMed), e.g., as disclosed in WO 2009/044273, incorporated by reference in its entirety.

TABLE 8 Amino acid sequences of other exemplary anti-LAG-3 antibody molecules BMS-986016 SEQ ID NO: 169 Heavy QVQLQQWGAGLLKPSETLSLTCAVYGGSFSDYYWNWIRQPPGKG chain LEWIGEINHRGSTNSNPSLKSRVTLSLDTSKNQFSLKLRSVTAADTA VYYCAFGYSDYEYNWFDPWGQGTLVTVSSASTKGPSVFPLAPCSR STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP CPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQF NWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRL TVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 170 Light EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLI chain YDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWP LTFGQGTNLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGEC IMP731 SEQ ID NO: 171 Heavy QVQLKESGPGLVAPSQSLSITCTVSGFSLTAYGVNWVRQPPGKGLE chain WLGMIWDDGSTDYNSALKSRLSISKDNSKSQVFLKMNSLQTDDTA RYYCAREGDVAFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLT CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 172 Light DIVMTQSPSSLAVSVGQKVTMSCKSSQSLLNGSNQKNYLAWYQQ chain KPGQSPKLLVYFASTRDSGVPDRFIGSGSGTDFTLTISSVQAEDLAD YFCLQHFGTPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC TIM-3 Inhibitors

In certain embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of TIM-3. In some embodiments, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with a TIM-3 inhibitor to treat a disease, e.g., cancer. In some embodiments, the TIM-3 inhibitor is MGB453 (Novartis) or TSR-022 (Tesaro).

Exemplary TIM-3 Inhibitors

In one embodiment, the TIM-3 inhibitor is an anti-TIM-3 antibody molecule. In one embodiment, the TIM-3 inhibitor is an anti-TIM-3 antibody molecule as disclosed in US 2015/0218274, published on Aug. 6, 2015, entitled “Antibody Molecules to TIM-3 and Uses Thereof,” incorporated by reference in its entirety.

In one embodiment, the anti-TIM-3 antibody molecule comprises at least one, two, three, four, five or six complementarity determining regions (CDRs) (or collectively all of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 9 (e.g., from the heavy and light chain variable region sequences of ABTIM3-hum11 or ABTIM3-hum03 disclosed in Table 9), or encoded by a nucleotide sequence shown in Table 9. In some embodiments, the CDRs are according to the Kabat definition (e.g., as set out in Table 9). In some embodiments, the CDRs are according to the Chothia definition (e.g., as set out in Table 9). In one embodiment, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions (e.g., conservative amino acid substitutions) or deletions, relative to an amino acid sequence shown in Table 9, or encoded by a nucleotide sequence shown in Table 9.

In one embodiment, the anti-TIM-3 antibody molecule comprises a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence of SEQ ID NO: 174, a VHCDR2 amino acid sequence of SEQ ID NO: 175, and a VHCDR3 amino acid sequence of SEQ ID NO: 176; and a light chain variable region (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 183, a VLCDR2 amino acid sequence of SEQ ID NO: 184, and a VLCDR3 amino acid sequence of SEQ ID NO: 185, each disclosed in Table 9. In one embodiment, the anti-TIM-3 antibody molecule comprises a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence of SEQ ID NO: 174, a VHCDR2 amino acid sequence of SEQ ID NO: 193, and a VHCDR3 amino acid sequence of SEQ ID NO: 176; and a light chain variable region (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 183, a VLCDR2 amino acid sequence of SEQ ID NO: 184, and a VLCDR3 amino acid sequence of SEQ ID NO: 185, each disclosed in Table 9.

In one embodiment, the anti-TIM-3 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 179, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 179. In one embodiment, the anti-TIM-3 antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 189, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 189. In one embodiment, the anti-TIM-3 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 195, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 195. In one embodiment, the anti-TIM-3 antibody molecule comprises a VL comprising the amino acid sequence of SEQ ID NO: 199, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 199. In one embodiment, the anti-TIM-3 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 179 and a VL comprising the amino acid sequence of SEQ ID NO: 189. In one embodiment, the anti-TIM-3 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 195 and a VL comprising the amino acid sequence of SEQ ID NO: 199.

In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 180, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 180. In one embodiment, the antibody molecule comprises a VL encoded by the nucleotide sequence of SEQ ID NO: 190, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 190. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 196, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 196. In one embodiment, the antibody molecule comprises a VL encoded by the nucleotide sequence of SEQ ID NO: 200, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 200. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 180 and a VL encoded by the nucleotide sequence of SEQ ID NO: 190. In one embodiment, the antibody molecule comprises a VH encoded by the nucleotide sequence of SEQ ID NO: 196 and a VL encoded by the nucleotide sequence of SEQ ID NO: 200.

In one embodiment, the anti-TIM-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 181, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 181. In one embodiment, the anti-TIM-3 antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 191, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 191. In one embodiment, the anti-TIM-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 197, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 197. In one embodiment, the anti-TIM-3 antibody molecule comprises a light chain comprising the amino acid sequence of SEQ ID NO: 201, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 201. In one embodiment, the anti-TIM-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 181 and a light chain comprising the amino acid sequence of SEQ ID NO: 191. In one embodiment, the anti-TIM-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 197 and a light chain comprising the amino acid sequence of SEQ ID NO: 201.

In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 182, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 182. In one embodiment, the antibody molecule comprises a light chain encoded by the nucleotide sequence of SEQ ID NO: 192, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 192. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 198, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 198. In one embodiment, the antibody molecule comprises a light chain encoded by the nucleotide sequence of SEQ ID NO: 202, or a nucleotide sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 202. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 182 and a light chain encoded by the nucleotide sequence of SEQ ID NO: 192. In one embodiment, the antibody molecule comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO: 198 and a light chain encoded by the nucleotide sequence of SEQ ID NO: 202.

The antibody molecules described herein can be made by vectors, host cells, and methods described in US 2015/0218274, incorporated by reference in its entirety.

TABLE 9 Amino acid and nucleotide sequences of exemplary anti-TIM-3 antibody molecules ABTIM3-hum11 SEQ ID NO: 174 HCDR1 SYNMH (Kabat) SEQ ID NO: 175 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 176 HCDR3 VGGAFPMDY (Kabat) SEQ ID NO: 177 HCDR1 GYTFTSY (Chothia) SEQ ID NO: 178 HCDR2 YPGNGD (Chothia) SEQ ID NO: 176 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: 179 VH QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQ GLEWMGDIYPGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLR SEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 180 DNA CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCC VH GGCTCTAGCGTGAAAGTTTCTTGTAAAGCTAGTGGCTACACCT TCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGA CACTAGTTATAATCAGAAGTTTAAGGGTAGAGTCACTATCACC GCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGG GCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGT GACCGTGTCTAGC SEQ ID NO: 181 Heavy QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQ chain GLEWMGDIYPGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLR SEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLA PCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKY GPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQ EDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS LG SEQ ID NO: 182 DNA CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCC heavy GGCTCTAGCGTGAAAGTTTCTTGTAAAGCTAGTGGCTACACCT chain TCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGA CACTAGTTATAATCAGAAGTTTAAGGGTAGAGTCACTATCACC GCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGG GCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGT GACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCC TCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGT GTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTC CCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGG TGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACAC TTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAA GCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCC GCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCAC CGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGT GACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGT GCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGC CAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAAC GGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCT AGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCC CGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAA ATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCT TCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCC AGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACT CAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAA GAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGAT GCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCC CTCTCCCTGGGA SEQ ID NO: 183 LCDR1 RASESVEYYGTSLMQ (Kabat) SEQ ID NO: 184 LCDR2 AASNVES (Kabat) SEQ ID NO: 185 LCDR3 QQSRKDPST (Kabat) SEQ ID NO: 186 LCDR1 SESVEYYGTSL (Chothia) SEQ ID NO: 187 LCDR2 AAS (Chothia) SEQ ID NO: 188 LCDR3 SRKDPS (Chothia) SEQ ID NO: 189 VL AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPG KAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYF CQQSRKDPSTFGGGTKVEIK SEQ ID NO: 190 DNA GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTG VL TGGGCGATAGAGTGACTATCACCTGTAGAGCTAGTGAATCAG TCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGA AGCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAA CGTGGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGT GGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGG ACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAG CACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: 191 Light AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPG chain KAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYF CQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 192 DNA GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTG light TGGGCGATAGAGTGACTATCACCTGTAGAGCTAGTGAATCAG chain TCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGA AGCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAA CGTGGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGT GGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGG ACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAG CACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGT GGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAAC TTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAAC GCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAG GACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACC CTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGC GAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGC TTCAACAGGGGCGAGTGC ABTIM3-hum03 SEQ ID NO: 174 HCDR1 SYNMH (Kabat) SEQ ID NO: 193 HCDR2 DIYPGQGDTSYNQKFKG (Kabat) SEQ ID NO: 176 HCDR3 VGGAFPMDY (Kabat) SEQ ID NO: 177 HCDR1 GYTFTSY (Chothia) SEQ ID NO: 194 HCDR2 YPGQGD (Chothia) SEQ ID NO: 176 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: 195 VH QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPG QGLEWIGDIYPGQGDTSYNQKFKGRATMTADKSTSTVYMELSSL RSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 196 DNA CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCC VH GGCGCTAGTGTGAAAGTTAGCTGTAAAGCTAGTGGCTATACTT TCACTTCTTATAATATGCACTGGGTCCGCCAGGCCCCAGGTCA AGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGA CACTTCCTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACC GCCGATAAGTCTACTTCTACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGG GCGGAGCCTTCCCAATGGACTACTGGGGTCAAGGCACCCTGG TCACCGTGTCTAGC SEQ ID NO: 197 Heavy QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPG chain QGLEWIGDIYPGQGDTSYNQKFKGRATMTADKSTSTVYMELSSL RSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPL APCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESK YGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS QEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQ EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLG SEQ ID NO: 198 DNA CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCC heavy GGCGCTAGTGTGAAAGTTAGCTGTAAAGCTAGTGGCTATACTT chain TCACTTCTTATAATATGCACTGGGTCCGCCAGGCCCCAGGTCA AGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGA CACTTCCTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACC GCCGATAAGTCTACTTCTACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGG GCGGAGCCTTCCCAATGGACTACTGGGGTCAAGGCACCCTGG TCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCC CCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCC CTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCG TGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCG GTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACA CTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCC CGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCA CCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAG TGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGG TGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACG CCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACC GCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAA CGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCC TAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCC CCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGA AATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGC TTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCC AGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACT CAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAA GAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGAT GCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCC CTCTCCCTGGGA SEQ ID NO: 183 LCDR1 RASESVEYYGTSLMQ (Kabat) SEQ ID NO: 184 LCDR2 AASNVES (Kabat) SEQ ID NO: 185 LCDR3 QQSRKDPST (Kabat) SEQ ID NO: 186 LCDR1 SESVEYYGTSL (Chothia) SEQ ID NO: 187 LCDR2 AAS (Chothia) SEQ ID NO: 188 LCDR3 SRKDPS (Chothia) SEQ ID NO: 199 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKP GQPPKLLIYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVY YCQQSRKDPSTFGGGTKVEIK SEQ ID NO: 200 DNA GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCC VL TGGGCGAGCGGGCTACTATTAACTGTAGAGCTAGTGAATCAG TCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGA AGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAA CGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAG TGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAG GACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTA GCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: 201 Light DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKP chain GQPPKLLIYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVY YCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 202 DNA GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCC light TGGGCGAGCGGGCTACTATTAACTGTAGAGCTAGTGAATCAG chain TCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGA AGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAA CGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAG TGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAG GACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTA GCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGG TGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCA GCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAA CTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAA CGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCA GGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGAC CCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTG CGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAG CTTCAACAGGGGCGAGTGC Other Exemplary TIM-3 Inhibitors

In one embodiment, the anti-TIM-3 antibody molecule is TSR-022 (AnaptysBio/Tesaro). In one embodiment, the anti-TIM-3 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of TSR-022. In one embodiment, the anti-TIM-3 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of APE5137 or APE5121, e.g., as disclosed in Table 10. APE5137, APE5121, and other anti-TIM-3 antibodies are disclosed in WO 2016/161270, incorporated by reference in its entirety.

In one embodiment, the anti-TIM-3 antibody molecule is the antibody clone F38-2E2. In one embodiment, the anti-TIM-3 antibody molecule comprises one or more of the CDR sequences (or collectively all of the CDR sequences), the heavy chain or light chain variable region sequence, or the heavy chain or light chain sequence of F38-2E2.

Further known anti-TIM-3 antibodies include those described, e.g., in WO 2016/111947, WO 2016/071448, WO 2016/144803, U.S. Pat. Nos. 8,552,156, 8,841,418, and 9,163,087, incorporated by reference in their entirety.

In one embodiment, the anti-TIM-3 antibody is an antibody that competes for binding with, and/or binds to the same epitope on TIM-3 as, one of the anti-TIM-3 antibodies described herein.

TABLE 10 Amino acid sequences of other exemplary anti-TIM-3 antibody molecules APE5137 SEQ ID NO: 203 VH EVQLLESGGGLVQPGGSLRLSCAAASGFTFSSYDMSWVRQAPGKGLD WVSTISGGGTYTYYQDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAV YYCASMDYWGQGTTVTVSSA SEQ ID NO: 204 VL DIQMTQSPSSLSASVGDRVTITCRASQSIRRYLNWYHQKPGKAPKLLI YGASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAVYYCQQSHSAPLT FGGGTKVEIKR APE5121 SEQ ID NO: 205 VH EVQVLESGGGLVQPGGSLRLYCVASGFTFSGSYAMSWVRQAPGKGL EWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAKKYYVGPADYWGQGTLVTVSSG SEQ ID NO: 206 VL DIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKNYLAWYQHKPG QPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQ QYYSSPLTFGGGTKIEVK Cytokines

In yet another embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more cytokines, including but not limited to, interferon, IL-2, IL-15, IL-7, or IL21. In certain embodiments, compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, are administered in combination with an IL-15/IL-15Ra complex. In some embodiments, the IL-15/IL-15Ra complex is selected from NIZ985 (Novartis), ATL-803 (Altor) or CYP0150 (Cytune).

Exemplary IL-15/IL-15Ra Complexes

In one embodiment, the cytokine is IL-15 complexed with a soluble form of IL-15 receptor alpha (IL-15Ra). The IL-15/IL-15Ra complex may comprise IL-15 covalently or noncovalently bound to a soluble form of IL-15Ra. In a particular embodiment, the human IL-15 is noncovalently bonded to a soluble form of IL-15Ra. In a particular embodiment, the human IL-15 of the formulation comprises an amino acid sequence of SEQ ID NO: 207 in Table 11 or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 207, and the soluble form of human IL-15Ra comprises an amino acid sequence of SEQ ID NO: 208 in Table 11, or an amino acid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 208, as described in WO 2014/066527, incorporated by reference in its entirety. The molecules described herein can be made by vectors, host cells, and methods described in WO 2007084342, incorporated by reference in its entirety.

TABLE 11 Amino acid and nucleotide sequences of exemplary IL-15/IL-15Ra complexes NIZ985 SEQ ID NO: 207 Human NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFL IL-15 LELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEE LEEKNIKEFLQSFVHIVQMFINTS SEQ ID NO: 208 Human ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTEC Soluble VLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSTVTTAGVTPQP IL-15Ra ESLSPSGKEPAASSPSSNNTAATTAAIVPGSQLMPSKSPSTGTTEI SSHESSHGTPSQTTAKNWELTASASHQPPGVYPQG Other Exemplar IL-15/IL-15Ra Complexes

In one embodiment, the IL-15/IL-15Ra complex is ALT-803, an IL-15/IL-15Ra Fc fusion protein (IL-15N72D:IL-15RaSu/Fc soluble complex). ALT-803 is described in WO 2008/143794, incorporated by reference in its entirety. In one embodiment, the IL-15/IL-15Ra Fc fusion protein comprises the sequences as disclosed in Table 12.

In one embodiment, the IL-15/IL-15Ra complex comprises IL-15 fused to the sushi domain of IL-15Ra (CYP0150, Cytune). The sushi domain of IL-15Ra refers to a domain beginning at the first cysteine residue after the signal peptide of IL-15Ra, and ending at the fourth cysteine residue after said signal peptide. The complex of IL-15 fused to the sushi domain of IL-15Ra is described in WO 2007/04606 and WO 2012/175222, incorporated by reference in their entirety. In one embodiment, the IL-15/IL-15Ra sushi domain fusion comprises the sequences as disclosed in Table 12.

TABLE 12 Amino acid sequences of other exemplary IL-15/IL-15Ra complexes ALT-803 SEQ ID NO: IL-15N72D NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCF 209 LLELQVISLESGDASIHDTVENLIILANDSLSSNGNVTESGCKEC EELEEKNIKEFLQSFVHIVQMFINTS SEQ ID NO: IL-15RaSu/Fc ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTE 210 CVLNKATNVAHWTTPSLKCIREPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSL TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK IL-15/IL-15Ra sushi domain fusion (CYP0150) SEQ ID NO: Human IL-15 NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCF 211 LLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKEC EELEXKNIKEFLQSFVHIVQMFINTS Where X is E or K SEQ ID NO: Human IL- ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTE 212 15Ra sushi CVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP and hinge domains

In yet another embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more agonists of toll like receptors (TLRs, e.g., TLR7, TLR8, TLR9) to treat a disease, e.g., cancer. In some embodiments, a compound of the present disclosure can be used in combination with a TLR7 agonist or a TLR7 agonist conjugate.

In some embodiments, the TLR7 agonist comprises a compound disclosed in International Application Publication No. WO2011/049677, which is hereby incorporated by reference in its entirety. In some embodiments, the TLR7 agonist comprises 3-(5-amino-2-(4-(2-(3,3-difluoro-3-phosphonopropoxy)ethoxy)-2-methylphenethyl)benzo[f][1,7]naphthyridin-8-yl)propanoic acid. In some embodiments, the TLR7 agonist comprises a compound of formula:

In another embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more angiogenesis inhibitors to treat cancer, e.g., Bevacizumab (Avastin®), axitinib (Inlyta®); Brivanib alaninate (BMS-582664, (S)—((R)-1-(4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[2,1-J][1,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate); Sorafenib (Nexavar®); Pazopanib (Votrient®); Sunitinib malate (Sutent®); Cediranib (AZD2171, CAS 288383-20-1); Vargatef (BIBF1120, CAS 928326-83-4); Foretinib (GSK1363089); Telatinib (BAY57-9352, CAS 332012-40-5); Apatinib (YN968D1, CAS 811803-05-1); Imatinib (Gleevec®); Ponatinib (AP24534, CAS 943319-70-8); Tivozanib (AV951, CAS 475108-18-0); Regorafenib (BAY73-4506, CAS 755037-03-7); Vatalanib dihydrochloride (PTK787, CAS 212141-51-0); Brivanib (BMS-540215, CAS 649735-46-6); Vandetanib (Caprelsa® or AZD6474); Motesanib diphosphate (AMG706, CAS 857876-30-3, N-(2,3-dihydro-3,3-dimethyl-1H-indol-6-yl)-2-[(4-pyridinylmethyl)amino]-3-pyridinecarboxamide, described in PCT Publication No. WO 02/066470); Dovitinib dilactic acid (TKI258, CAS 852433-84-2); Linfanib (ABT869, CAS 796967-16-3); Cabozantinib (XL184, CAS 849217-68-1); Lestaurtinib (CAS 111358-88-4); N-[5-[[[5-(1,1-Dimethylethyl)-2-oxazolyl]methyl]thio]-2-thiazolyl]-4-piperidinecarboxamide (BMS38703, CAS 345627-80-7); (3R,4R)-4-Amino-1-((4-((3-methoxyphenyl)amino)pyrrolo[2,1-f][1,2,4]triazin-5-yl)methyl)piperidin-3-ol (BMS690514); N-(3,4-Dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aa,5(3,6aa)-octahydro-2-methylcyclopenta[c]pyrrol-5-yl]methoxy]-4-quinazolinamine (XL647, CAS 781613-23-8); 4-Methyl-3-[[1-methyl-6-(3-pyridinyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl]amino]-N-[3-(trifluoromethyl)phenyl]-benzamide (BHG712, CAS 940310-85-0); or Aflibercept (Eylea®).

In another embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more heat shock protein inhibitors to treat cancer, e.g., Tanespimycin (17-allylamino-17-demethoxygeldanamycin, also known as KOS-953 and 17-AAG, available from SIGMA, and described in U.S. Pat. No. 4,261,989); Retaspimycin (IPI504), Ganetespib (STA-9090); [6-Chloro-9-(4-methoxy-3,5-dimethylpyridin-2-ylmethyl)-9H-purin-2-yl]amine (BIIB021 or —CNF2024, CAS 848695-25-0); trans-4-[[2-(Aminocarbonyl)-5-[4,5,6,7-tetrahydro-6,6-dimethyl-4-oxo-3-(trifluoromethyl)-1H-indazol-1-yl]phenyl]amino]cyclohexyl glycine ester (SNX5422 or PF04929113, CAS 908115-27-5); 5-[2,4-Dihydroxy-5-(1-methylethyl)phenyl]-N-ethyl-4-[4-(4-morpholinylmethyl)phenyl]-3-Isoxazolecarboxamide (AUY922, CAS 747412-49-3); or 17-Dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG).

In yet another embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more HDAC inhibitors or other epigenetic modifiers. Exemplary HDAC inhibitors include, but not limited to, Voninostat (Zolinza®); Romidepsin (Istodax®); Treichostatin A (TSA); Oxamflatin; Vorinostat (Zolinza®, Suberoylanilide hydroxamic acid); Pyroxamide (syberoyl-3-aminopyridineamide hydroxamic acid); Trapoxin A (RF-1023A); Trapoxin B (RF-10238); Cyclo[(αS,2S)-α-amino-η-oxo-2-oxiraneoctanoyl-O-methyl-D-tyrosyl-L-isoleucyl-L-prolyl] (Cyl-1); Cyclo[(αS,2S)-α-amino-η-oxo-2-oxiraneoctanoyl-O-methyl-D-tyrosyl-L-isoleucyl-(2S)-2-piperidinecarbonyl] (Cyl-2); Cyclic[L-alanyl-D-alanyl-(2S)-η-oxo-L-α-aminooxiraneoctanoyl-D-prolyl] (HC-toxin); Cyclo[(αS,2S)-α-amino-η-oxo-2-oxiraneoctanoyl-D-phenylalanyl-L-leucyl-(2S)-2-piperidinecarbonyl] (WF-3161); Chlamydocin ((S)-Cyclic(2-methylalanyl-L-phenylalanyl-D-prolyl-η-oxo-L-α-aminooxiraneoctanoyl); Apicidin (Cyclo(8-oxo-L-2-aminodecanoyl-1-methoxy-L-tryptophyl-L-isoleucyl-D-2-piperidinecarbonyl); Romidepsin (Istodax®, FR-901228); 4-Phenylbutyrate; Spiruchostatin A; Mylproin (Valproic acid); Entinostat (MS-275, N-(2-Aminophenyl)-4-[N-(pyridine-3-yl-methoxycarbonyl)-amino-methyl]-benzamide); Depudecin (4,5:8,9-dianhydro-1,2,6,7,11-pentadeoxy-D-threo-D-ido-Undeca-1,6-dienitol); 4-(Acetylamino)-N-(2-aminophenyl)-benzamide (also known as CI-994); N1-(2-Aminophenyl)-N8-phenyl-octanediamide (also known as BML-210); 4-(Dimethylamino)-N-(7-(hydroxyamino)-7-oxoheptyl)benzamide (also known as M344); (E)-3-(4-(((2-(1H-indol-3-yl)ethyl)(2-hydroxyethyl)amino)-methyl)phenyl)-N-hydroxyacrylamide; Panobinostat(Farydak®); Mocetinostat, and Belinostat (also known as PXD101, Beleodaq®, or (2E)-N-Hydroxy-3-[3-(phenylsulfamoyl)phenyl]prop-2-enamide), or chidamide (also known as CS055 or HBI-8000, (E)-N-(2-amino-5-fluorophenyl)-4-((3-(pyridin-3-yl)acrylamido)methyl)benzamide). Other epigenetic modifiers include but not limited to inhibitors of EZH2 (enhancer of zeste homolog 2), EED (embryonic ectoderm development), or LSD1 (lysine-specific histone demethylase 1A or KDM1A).

In yet another embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more inhibitors of indoleamine-pyrrole 2,3-dioxygenase (IDO), for example, Indoximod (also known as NLG-8189), α-Cyclohexyl-5H-imidazo[5,1-a]isoindole-5-ethanol (also known as NLG919), or (4E)-4-[(3-Chloro-4-fluoroanilino)-nitrosomethylidene]-1,2,5-oxadiazol-3-amine (also known as INCB024360), to treat cancer.

Chimeric Antigen Receptors

The present disclosure provides for the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof for use in combination with adoptive immunotherapy methods and reagents such as chimeric antigen receptor (CAR) immune effector cells, e.g., T cells, or chimeric TCR-transduced immune effector cells, e.g., T cells. This section describes CAR technology generally that is useful in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, and describes CAR reagents, e.g., cells and compositions, and methods.

In general, aspects of the present disclosure pertain to or include an isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen binding domain (e.g., antibody or antibody fragment, TCR or TCR fragment) that binds to a tumor antigen as described herein, a transmembrane domain (e.g., a transmembrane domain described herein), and an intracellular signalling domain (e.g., an intracellular signalling domain described herein) (e.g., an intracellular signalling domain comprising a costimulatory domain (e.g., a costimulatory domain described herein) and/or a primary signalling domain (e.g., a primary signalling domain described herein). In other aspects, the present disclosure includes: host cells containing the above nucleic acids and isolated proteins encoded by such nucleic acid molecules. CAR nucleic acid constructs, encoded proteins, containing vectors, host cells, pharmaceutical compositions, and methods of administration and treatment related to the present disclosure are disclosed in detail in International Patent Application Publication No. WO2015142675, which is incorporated by reference in its entirety.

In one aspect, the disclosure pertains to an isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen binding domain (e.g., antibody or antibody fragment, TCR or TCR fragment) that binds to a tumor-supporting antigen (e.g., a tumor-supporting antigen as described herein), a transmembrane domain (e.g., a transmembrane domain described herein), and an intracellular signalling domain (e.g., an intracellular signalling domain described herein) (e.g., an intracellular signalling domain comprising a costimulatory domain (e.g., a costimulatory domain described herein) and/or a primary signalling domain (e.g., a primary signalling domain described herein). In some embodiments, the tumor-supporting antigen is an antigen present on a stromal cell or a myeloid-derived suppressor cell (MDSC). In other aspects, the disclosure features polypeptides encoded by such nucleic acids and host cells containing such nucleic acids and/or polypeptides.

Alternatively, aspects of the disclosure pertain to isolated nucleic acid encoding a chimeric T cell receptor (TCR) comprising a TCR alpha and/or TCR beta variable domain with specificity for a cancer antigen described herein. See for example, Dembic et al., Nature, 320, 232-238 (1986), Schumacher, Nat. Rev. Immunol., 2, 512-519 (2002), Kershaw et al., Nat. Rev. Immunol., 5, 928-940 (2005), Xue et al., Clin. Exp. Immunol., 139, 167-172 (2005), Rossig et al., Mol. Ther., 10, 5-18 (2004), and Murphy et al., Immunity, 22, 403-414 (2005); (Morgan et al. J. Immunol., 171, 3287-3295 (2003), Hughes et al., Hum. Gene Ther., 16, 1-16 (2005), Zhao et al., J. Immunol., 174, 4415-4423 (2005), Roszkowski et al., Cancer Res., 65, 1570-1576 (2005), and Engels et al., Hum. Gene Ther., 16, 799-810 (2005); US2009/03046557, the contents of which are hereby incorporated by reference in their entirety. Such chimeric TCRs may recognize, for example, cancer antigens such as MART-1, gp-100, p53, and NY-ESO-1, MAGE A3/A6, MAGEA3, SSX2, HPV-16 E6 or HPV-16 E7. In other aspects, the disclosure features polypeptides encoded by such nucleic acids and host cells containing such nucleic acids and/or polypeptides.

Sequences of non-limiting examples of various components that can be part of a CAR are listed in Table 11a, where “aa” stands for amino acids, and “na” stands for nucleic acids that encode the corresponding peptide.

TABLE 11a Sequences of various components of CAR (aa—amino acid sequence, na—nucleic acid sequence). SEQ ID NO: description Sequence SEQ ID EF-1 CGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGC NO: 270 promoter CCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAA (na) CCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTG ATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGA ACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGC AACGGGTTTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGT TCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGC CTTGAATTACTTCCACCTGGCTGCAGTACGTGATTCTTGATCCC GAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGC GCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGC CTGGGCGCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCG CGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATT TTTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTT GTAAATGCGGGCCAAGATCTGCACACTGGTATTTCGGTTTTTG GGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACAT GTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCG GACGGGGGTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGG CCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTG GCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTT CCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACGCGGCGC TCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAAG GGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGGAG TACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTT GGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGAT GGAGTTTCCCCACACTGAGTGGGTGGAGACTGAAGTTAGGCC AGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTG AGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCA AAGTTTTTTTCTTCCATTTCAGGTGTCGTGA SEQ ID Leader (aa) MALPVTALLLPLALLLHAARP NO: 268 SEQ ID Leader (na) ATGGCCCTGCCTGTGACAGCCCTGCTGCTGCCTCTGGCTCTGC NO: TGCTGCATGCCGCTAGACCC 287 SEQ ID Leader (na) ATGGCCCTCCCTGTCACCGCCCTGCTGCTTCCGCTGGCTCTTCT NO: GCTCCACGCCGCTCGGCCC 288 SEQ ID CD8 hinge TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD NO: 250 (aa) SEQ ID CD8 hinge ACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACC NO: 254 (na) ATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGC CAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCG CCTGTGAT SEQ ID IgG4 hinge ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVV NO: 253 (aa) DVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTL PPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQK SLSLSLGKM SEQ ID IgG4 hinge GAGAGCAAGTACGGCCCTCCCTGCCCCCCTTGCCCTGCCCCCG NO: 255 (na) AGTTCCTGGGCGGACCCAGCGTGTTCCTGTTCCCCCCCAAGCC CAAGGACACCCTGATGATCAGCCGGACCCCCGAGGTGACCTG TGTGGTGGTGGACGTGTCCCAGGAGGACCCCGAGGTCCAGTT CAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGAC CAAGCCCCGGGAGGAGCAGTTCAATAGCACCTACCGGGTGGT GTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAA GGAATACAAGTGTAAGGTGTCCAACAAGGGCCTGCCCAGCAG CATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCTCGGGA GCCCCAGGTGTACACCCTGCCCCCTAGCCAAGAGGAGATGAC CAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTAC CCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCC GAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGAC GGCAGCTTCTTCCTGTACAGCCGGCTGACCGTGGACAAGAGCC GGTGGCAGGAGGGCAACGTCTTTAGCTGCTCCGTGATGCACG AGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGT CCCTGGGCAAGATG SEQ ID IgD hinge RWPESPKAQASSVPTAQPQAEGSLAKATTAPATTRNTGRGGEEK NO: 256 (aa) KKEKEKEEQEERETKTPECPSHTQPLGVYLLTPAVQDLWLRDKA TFTCFVVGSDLKDAHLTWEVAGKVPTGGVEEGLLERHSNGSQSQ HSRLTLPRSLWNAGTSVTCTLNHPSLPPQRLMALREPAAQAPVK LSLNLLASSDPPEAASWLLCEVSGFSPPNILLMWLEDQREVNTSG FAPARPPPQPGSTTFWAWSVLRVPAPPSPQPATYTCVVSHEDSRT LLNASRSLEVSYVTDH SEQ ID IgD hinge AGGTGGCCCGAAAGTCCCAAGGCCCAGGCATCTAGTGTTCCT NO: 257 (na) ACTGCACAGCCCCAGGCAGAAGGCAGCCTAGCCAAAGCTACT ACTGCACCTGCCACTACGCGCAATACTGGCCGTGGCGGGGAG GAGAAGAAAAAGGAGAAAGAGAAAGAAGAACAGGAAGAGA GGGAGACCAAGACCCCTGAATGTCCATCCCATACCCAGCCGC TGGGCGTCTATCTCTTGACTCCCGCAGTACAGGACTTGTGGCT TAGAGATAAGGCCACCTTTACATGTTTCGTCGTGGGCTCTGAC CTGAAGGATGCCCATTTGACTTGGGAGGTTGCCGGAAAGGTA CCCACAGGGGGGGTTGAGGAAGGGTTGCTGGAGCGCCATTCC AATGGCTCTCAGAGCCAGCACTCAAGACTCACCCTTCCGAGAT CCCTGTGGAACGCCGGGACCTCTGTCACATGTACTCTAAATCA TCCTAGCCTGCCCCCACAGCGTCTGATGGCCCTTAGAGAGCCA GCCGCCCAGGCACCAGTTAAGCTTAGCCTGAATCTGCTCGCCA GTAGTGATCCCCCAGAGGCCGCCAGCTGGCTCTTATGCGAAGT GTCCGGCTTTAGCCCGCCCAACATCTTGCTCATGTGGCTGGAG GACCAGCGAGAAGTGAACACCAGCGGCTTCGCTCCAGCCCGG CCCCCACCCCAGCCGGGTTCTACCACATTCTGGGCCTGGAGTG TCTTAAGGGTCCCAGCACCACCTAGCCCCCAGCCAGCCACATA CACCTGTGTTGTGTCCCATGAAGATAGCAGGACCCTGCTAAAT GCTTCTAGGAGTCTGGAGGTTTCCTACGTGACTGACCATT SEQ ID GS GGGGSGGGGS NO: 258 hinge/linker (aa) SEQ ID GS GGTGGCGGAGGTTCTGGAGGTGGAGGTTCC NO: 259 hinge/linker (na) SEQ ID CD8 IYIWAPLAGTCGVLLLSLVITLYC NO: 251 transmembrane (aa) SEQ ID CD8 ATCTACATCTGGGCGCCCTTGGCCGGGACTTGTGGGGTCCTTC NO: 252 transmembrane TCCTGTCACTGGTTATCACCCTTTACTGC (na) SEQ ID CD8 ATCTACATTTGGGCCCCTCTGGCTGGTACTTGCGGGGTCCTGC NO: 289 transmembrane TGCTTTCACTCGTGATCACTCTTTACTGT (na) SEQ ID 4-1BB KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL NO: 264 intracellular domain (aa) SEQ ID 4-1BB AAACGGGGCAGAAAGAAACTCCTGTATATATTCAAACAACCA NO: 266 intracellular TTTATGAGACCAGTACAAACTACTCAAGAGGAAGATGGCTGT domain (na) AGCTGCCGATTTCCAGAAGAAGAAGAAGGAGGATGTGAACTG SEQ ID 4-1BB AAGCGCGGTCGGAAGAAGCTGCTGTACATCTTTAAGCAACCC NO: 290 intracellular TTCATGAGGCCTGTGCAGACTACTCAAGAGGAGGACGGCTGT domain (na) TCATGCCGGTTCCCAGAGGAGGAGGAAGGCGGCTGCGAACTG SEQ ID CD27 (aa) QRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPAC NO: 265 SP SEQ ID CD27 (na) Caacgaaggaaatatagatcaaacaaaggagaaagtcctgtggagcctgcagagccttgtcgttaca NO: 267 gctgccccagggaggaggagggcagcaccatccccatccaggaggattaccgaaaaccggagcct gcctgctccccc SEQ ID CD3-zeta RVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDP NO: 260 (aa) EMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKG HDGLYQGLSTATKDTYDALHMQALPPR SEQ ID CD3-zeta AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACAAG NO: 262 (na) CAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGA AGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGAC CCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGGA AGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGC CTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAA GGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAA GGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGC SEQ ID CD3-zeta CGCGTGAAATTCAGCCGCAGCGCAGATGCTCCAGCCTACAAG NO: 291 (na) CAGGGGCAGAACCAGCTCTACAACGAACTCAATCTTGGTCGG AGAGAGGAGTACGACGTGCTGGACAAGCGGAGAGGACGGGA CCCAGAAATGGGCGGGAAGCCGCGCAGAAAGAATCCCCAAG AGGGCCTGTACAACGAGCTCCAAAAGGATAAGATGGCAGAAG CCTATAGCGAGATTGGTATGAAAGGGGAACGCAGAAGAGGCA AAGGCCACGACGGACTGTACCAGGGACTCAGCACCGCCACCA AGGACACCTATGACGCTCTTCACATGCAGGCCCTGCCGCCTCGG SEQ ID CD3-zeta RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDP NO: 261 (aa) EMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKG HDGLYQGLSTATKDTYDALHMQALPPR SEQ ID CD3-zeta AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAG NO: 263 (na) CAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGA AGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGAC CCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGGA AGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGC CTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAA GGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAA GGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGC SEQ ID Linker (aa) GGGGS NO: 292 SEQ ID PD-1 Pgwfldspdrpwnpptfspallvvtegdnatftcsfsntsesfvlnwyrmspsnqtdklaafpedrs NO: 293 extracellular qpgqdcrfrvtqlpngrdfhmsvvrarrndsgtylcgaislapkaqikeslraelrvterraevptahp domain (aa) spsprpagqfqtlv SEQ ID PD-1 Cccggatggtttctggactctccggatcgcccgtggaatcccccaaccttctcaccggcactcttggttg NO: 294 extracellular tgactgagggcgataatgcgaccttcacgtgctcgttctccaacacctccgaatcattcgtgctgaactg domain (na) gtaccgcatgagcccgtcaaaccagaccgacaagctcgccgcgtttccggaagatcggtcgcaaccg ggacaggattgtcggttccgcgtgactcaactgccgaatggcagagacttccacatgagcgtggtccg cgctaggcgaaacgactccgggacctacctgtgcggagccatctcgctggcgcctaaggcccaaatc aaagagagcttgagggccgaactgagagtgaccgagcgcagagctgaggtgccaactgcacatcca tccccatcgcctcggcctgcggggcagtttcagaccctggtc SEQ ID PD-1 CAR Malpvtalllplalllhaarppgwfldspdrpwnpptfspallvvtegdnatftcsfsntsesfvlnwy NO: 295 (aa) with rmspsnqtdklaafpedrsqpgqdcrfrvtqlpngrdfhmsvvrarrndsgtylcgaislapkaqik signal eslraelrvterraevptahpspsprpagqfqtlvtttpaprpptpaptiasqplslrpeacrpaaggavh trgldfacdiyiwaplagtcgvlllslvitlyckrgrkkllyifkqpfmrpvqttqeedgcscrfpeeee ggcelrvkfsrsadapaykqgqnqlynelnlgrreeydvldkrrgrdpemggkprrknpqeglyn elqkdkmaeayseigmkgerrrgkghdglyqglstatkdtydalhmqalppr SEQ ID PD-1 CAR Atggccctccctgtcactgccctgcttctccccctcgcactcctgctccacgccgctagaccacccgga NO: 296 (na) tggtttctggactctccggatcgcccgtggaatcccccaaccttctcaccggcactcttggttgtgactga gggcgataatgcgaccttcacgtgctcgttctccaacacctccgaatcattcgtgctgaactggtaccgc atgagcccgtcaaaccagaccgacaagctcgccgcgtttccggaagatcggtcgcaaccgggacag gattgtcggttccgcgtgactcaactgccgaatggcagagacttccacatgagcgtggtccgcgctagg cgaaacgactccgggacctacctgtgcggagccatctcgctggcgcctaaggcccaaatcaaagaga gcttgagggccgaactgagagtgaccgagcgcagagctgaggtgccaactgcacatccatccccatc gcctcggcctgcggggcagtttcagaccctggtcacgaccactccggcgccgcgcccaccgactccg gccccaactatcgcgagccagcccctgtcgctgaggccggaagcatgccgccctgccgccggaggt gctgtgcatacccggggattggacttcgcatgcgacatctacatttgggctcctctcgccggaacttgtg gcgtgctccttctgtccctggtcatcaccctgtactgcaagcggggtcggaaaaagcttctgtacattttc aagcagcccttcatgaggcccgtgcaaaccacccaggaggaggacggttgctcctgccggttccccg aagaggaagaaggaggttgcgagctgcgcgtgaagttctcccggagcgccgacgcccccgcctata agcagggccagaaccagctgtacaacgaactgaacctgggacggcgggaagagtacgatgtgctgg acaagcggcgcggccgggaccccgaaatgggcgggaagcctagaagaaagaaccctcaggaagg cctgtataacgagctgcagaaggacaagatggccgaggcctactccgaaattgggatgaagggagag cggcggaggggaaaggggcacgacggcctgtaccaaggactgtccaccgccaccaaggacacata cgatgccctgcacatgcaggcccttccccctcgc SEQ ID Linker (aa) (Gly-Gly-Gly-Ser)n, where n = 1-10 NO: 297 SEQ ID Linker (aa) (Gly4 Ser)4 NO: 215 SEQ ID Linker (aa) (Gly4 Ser)3 NO: 216 SEQ ID Linker (aa) (Gly3Ser) NO: 297 SEQ ID polyA (na) [a]₅₀₋₅₀₀₀ NO: 298 SEQ ID PD1 CAR Pgwfldspdrpwnpptfspallvvtegdnatftcsfsntsesfvlnwvrmspsnqtdklaafpedrs NO: 299 (aa) qpgqdcrfrvtqlpngrdfhmsvvrarrndsgtylcgaislapkaqikeslraelryterraevptahp spsprpagqfqtlvtttpaprpptpaptiasqplslrpeacrpaaggavhtrgldfacdiyiwaplagtc gvlllslvitlyckrgrkkllyifkqpfmrpvqttqeedgcscrfpeeeeggcelrvkfsrsadapayk qgqnqlynelnlgrreeydvldkrrgrdpemggkprrknpqeglynelqkdkmaeayseigmk gerrrgkghdglyqglstatkdtydalhmqalppr SEQ ID ICOS TKKKYSSSVHDPNGEYMFMRAVNTAKKSRLTDVTL NO: 300 intracellular domain (aa) SEQ ID ICOS ACAAAAAAGAAGTATTCATCCAGTGTGCACGACCCTAACGGT NO: 301 intracellular GAATACATGTTCATGAGAGCAGTGAACACAGCCAAAAAATCC domain (na) AGACTCACAGATGTGACCCTA SEQ ID ICOS TM TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDF NO: 302 domain (aa) WLPIGCAAFVVVCILGCILICWL SEQ ID ICOS TM ACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACC NO: 303 domain (na) ATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGC CAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCG CCTGTGATTTCTGGTTACCCATAGGATGTGCAGCCTTTGTTGTA GTCTGCATTTTGGGATGCATACTTATTTGTTGGCTT SEQ ID CD28 RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS NO: 304 intracellular domain (aa) SEQ ID CD28 AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAAC NO: 305 intracellular ATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGC domain (na) CCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCC Targets

The present disclosure provides cells, e.g., immune effector cells (e.g., T cells, NK cells), that comprise or at any time comprised a gRNA molecule or CRISPR system as described herein, that are further engineered to contain one or more CARs that direct the immune effector cells to undesired cells (e.g., cancer cells). This is achieved through an antigen binding domain on the CAR that is specific for a cancer associated antigen. There are two classes of cancer associated antigens (tumor antigens) that can be targeted by the CARs of the instant disclosure: (1) cancer associated antigens that are expressed on the surface of cancer cells; and (2) cancer associated antigens that itself is intracellular, however, a fragment of such antigen (peptide) is presented on the surface of the cancer cells by MHC (major histocompatibility complex).

In some embodiments, the tumor antigen is chosen from one or more of: CD19; CD123; CD22; CD30; CD171; CS-1 (also referred to as CD2 subset 1, CRACC, SLAMF7, CD319, and 19A24); C-type lectin-like molecule-1 (CLL-1 or CLECL1); CD33; epidermal growth factor receptor variant III (EGFRvIII); ganglioside G2 (GD2); ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer); TNF receptor family member B cell maturation (BCMA); Tn antigen ((Tn Ag) or (GalNAca-Ser/Thr)); prostate-specific membrane antigen (PSMA); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Fms-Like Tyrosine Kinase 3 (FLT3); Tumor-associated glycoprotein 72 (TAG72); CD38; CD44v6; Carcinoembryonic antigen (CEA); Epithelial cell adhesion molecule (EPCAM); B7H3 (CD276); KIT (CD117); Interleukin-13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2); Mesothelin; Interleukin 11 receptor alpha (IL-11Ra); prostate stem cell antigen (PSCA); Protease Serine 21 (Testisin or PRSS21); vascular endothelial growth factor receptor 2 (VEGFR2); Lewis(Y) antigen; CD24; Platelet-derived growth factor receptor beta (PDGFR-beta); Stage-specific embryonic antigen-4 (SSEA-4); CD20; Folate receptor alpha; Receptor tyrosine-protein kinase ERBB2 (Her2/neu); Mucin 1, cell surface associated (MUC1); epidermal growth factor receptor (EGFR); neural cell adhesion molecule (NCAM); Prostase; prostatic acid phosphatase (PAP); elongation factor 2 mutated (ELF2M); Ephrin B2; fibroblast activation protein alpha (FAP); insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX); Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2); glycoprotein 100 (gp100); oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl); tyrosinase; ephrin type-A receptor 2 (EphA2); Fucosyl GM1; sialyl Lewis adhesion molecule (sLe); ganglioside GM3 (aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer); transglutaminase 5 (TGS5); high molecular weight-melanoma-associated antigen (HMWMAA); o-acetyl-GD2 ganglioside (OAcGD2); Folate receptor beta; tumor endothelial marker 1 (TEM1/CD248); tumor endothelial marker 7-related (TEM7R); claudin 6 (CLDN6); thyroid stimulating hormone receptor (TSHR); G protein-coupled receptor class C group 5, member D (GPRC5D); chromosome X open reading frame 61 (CXORF61); CD97; CD179a; anaplastic lymphoma kinase (ALK); Polysialic acid; placenta-specific 1 (PLAC1); hexasaccharide portion of globoH glycoceramide (GloboH); mammary gland differentiation antigen (NY-BR-1); uroplakin 2 (UPK2); Hepatitis A virus cellular receptor 1 (HAVCR1); adrenoceptor beta 3 (ADRB3); pannexin 3 (PANX3); G protein-coupled receptor 20 (GPR20); lymphocyte antigen 6 complex, locus K 9 (LY6K); Olfactory receptor 51E2 (OR51E2); TCR Gamma Alternate Reading Frame Protein (TARP); Wilms tumor protein (WT1); Cancer/testis antigen 1 (NY-ESO-1); Cancer/testis antigen 2 (LAGE-1a); Melanoma-associated antigen 1 (MAGE-A1); ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML); sperm protein 17 (SPA17); X Antigen Family, Member 1A (XAGE1); angiopoietin-binding cell surface receptor 2 (Tie 2); melanoma cancer testis antigen-1 (MAD-CT-1); melanoma cancer testis antigen-2 (MAD-CT-2); Fos-related antigen 1; tumor protein p53 (p53); p53 mutant; prostein; surviving; telomerase; prostate carcinoma tumor antigen-1 (PCTA-1 or Galectin 8), melanoma antigen recognized by T cells 1 (MelanA or MART1); Rat sarcoma (Ras) mutant; human Telomerase reverse transcriptase (hTERT); sarcoma translocation breakpoints; melanoma inhibitor of apoptosis (ML-IAP); ERG (transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene); N-Acetyl glucosaminyl-transferase V (NA17); paired box protein Pax-3 (PAX3); Androgen receptor; Cyclin B1; v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN); Ras Homolog Family Member C (RhoC); Tyrosinase-related protein 2 (TRP-2); Cytochrome P450 1B1 (CYP1B1); CCCTC-Binding Factor (Zinc Finger Protein)-Like (BORIS or Brother of the Regulator of Imprinted Sites), Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3); Paired box protein Pax-5 (PAX5); proacrosin binding protein sp32 (OY-TES1); lymphocyte-specific protein tyrosine kinase (LCK); A kinase anchor protein 4 (AKAP-4); synovial sarcoma, X breakpoint 2 (SSX2); Receptor for Advanced Glycation Endproducts (RAGE-1); renal ubiquitous 1 (RU1); renal ubiquitous 2 (RU2); legumain; human papilloma virus E6 (HPV E6); human papilloma virus E7 (HPV E7); intestinal carboxyl esterase; heat shock protein 70-2 mutated (mut hsp70-2); CD79a; CD79b; CD72; Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); Fc fragment of IgA receptor (FCAR or CD89); Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); CD300 molecule-like family member f (CD300LF); C-type lectin domain family 12 member A (CLEC12A); bone marrow stromal cell antigen 2 (BST2); EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2); lymphocyte antigen 75 (LY75); Glypican-3 (GPC3); Fc receptor-like 5 (FCRL5); and immunoglobulin lambda-like polypeptide 1 (IGLL1).

A CAR described herein can comprise an antigen binding domain (e.g., antibody or antibody fragment, TCR or TCR fragment) that binds to a tumor-supporting antigen (e.g., a tumor-supporting antigen as described herein). In some embodiments, the tumor-supporting antigen is an antigen present on a stromal cell or a myeloid-derived suppressor cell (MDSC). Stromal cells can secrete growth factors to promote cell division in the microenvironment. MDSC cells can inhibit T cell proliferation and activation. Without wishing to be bound by theory, in some embodiments, the CAR-expressing cells destroy the tumor-supporting cells, thereby indirectly inhibiting tumor growth or survival.

In embodiments, the stromal cell antigen is chosen from one or more of: bone marrow stromal cell antigen 2 (BST2), fibroblast activation protein (FAP) and tenascin. In an embodiment, the FAP-specific antibody is, competes for binding with, or has the same CDRs as, sibrotuzumab. In embodiments, the MDSC antigen is chosen from one or more of: CD33, CD11b, C14, CD15, and CD66b. Accordingly, in some embodiments, the tumor-supporting antigen is chosen from one or more of: bone marrow stromal cell antigen 2 (BST2), fibroblast activation protein (FAP) or tenascin, CD33, CD1 b, C14, CD15, and CD66b. Antigen Binding Domain Structures

In some embodiments, the antigen binding domain of the encoded CAR molecule comprises an antibody, an antibody fragment, an scFv, a Fv, a Fab, a (Fab′)2, a single domain antibody (SDAB), a VH or VL domain, a camelid VHH domain or a bi-functional (e.g. bi-specific) hybrid antibody (e.g., Lanzavecchia et al., Eur. J. Immunol. 17, 105 (1987)).

In some instances, scFvs can be prepared according to method known in the art (see, for example, Bird et al., (1988) Science 242:423-426 and Huston et al., (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). ScFv molecules can be produced by linking VH and VL regions together using flexible polypeptide linkers. The scFv molecules comprise a linker (e.g., a Ser-Gly linker) with an optimized length and/or amino acid composition. The linker length can greatly affect how the variable regions of a scFv fold and interact. In fact, if a short polypeptide linker is employed (e.g., between 5-10 amino acids) intrachain folding is prevented. Interchain folding is also required to bring the two variable regions together to form a functional epitope binding site. For examples of linker orientation and size see, e.g., Hollinger et al. 1993 Proc Natl Acad. Sci. U.S.A. 90:6444-6448, U.S. Patent Application Publication Nos. 2005/0100543, 2005/0175606, 2007/0014794, and PCT publication Nos. WO2006/020258 and WO2007/024715, is incorporated herein by reference.

An scFv can comprise a linker of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, or more amino acid residues between its VL and VH regions. The linker sequence may comprise any naturally occurring amino acid. In some embodiments, the linker sequence comprises amino acids glycine and serine. In another embodiment, the linker sequence comprises sets of glycine and serine repeats such as (Gly₄Ser)n, where n is a positive integer equal to or greater than 1 (SEQ ID NO: 217). In one embodiment, the linker can be (Gly₄Ser)₄ (SEQ ID NO: 215) or (Gly₄Ser)₃ (SEQ ID NO: 216). Variation in the linker length may retain or enhance activity, giving rise to superior efficacy in activity studies.

In another aspect, the antigen binding domain is a T cell receptor (“TCR”), or a fragment thereof, for example, a single chain TCR (scTCR). Methods to make such TCRs are known in the art. See, e.g., Willemsen R A et al, Gene Therapy 7: 1369-1377 (2000); Zhang T et al, Cancer Gene Ther 11: 487-496 (2004); Aggen et al, Gene Ther. 19(4):365-74 (2012) (references are incorporated herein by its entirety). For example, scTCR can be engineered that contains the Vα and Vβ genes from a T cell clone linked by a linker (e.g., a flexible peptide). This approach is very useful to cancer associated target that itself is intracellular, however, a fragment of such antigen (peptide) is presented on the surface of the cancer cells by MHC.

In certain embodiments, the encoded antigen binding domain has a binding affinity KD of 10⁻⁴ M to 10⁻⁸ M.

In one embodiment, the encoded CAR molecule comprises an antigen binding domain that has a binding affinity KD of 10⁻⁴ M to 10⁻⁸ M, e.g., 10⁻⁵ M to 10⁻⁷ M, e.g., 10⁻⁶ M or 10⁻⁷ M, for the target antigen. In one embodiment, the antigen binding domain has a binding affinity that is at least five-fold, 10-fold, 20-fold, 30-fold, 50-fold, 100-fold or 1,000-fold less than a reference antibody, e.g., an antibody described herein. In one embodiment, the encoded antigen binding domain has a binding affinity at least 5-fold less than a reference antibody (e.g., an antibody from which the antigen binding domain is derived). In one aspect such antibody fragments are functional in that they provide a biological response that can include, but is not limited to, activation of an immune response, inhibition of signal-transduction origination from its target antigen, inhibition of kinase activity, and the like, as will be understood by a skilled artisan. In one aspect, the antigen binding domain of the CAR is a scFv antibody fragment that is humanized compared to the murine sequence of the scFv from which it is derived.

In one aspect, the antigen binding domain of a CAR of the disclosure (e.g., a scFv) is encoded by a nucleic acid molecule whose sequence has been codon optimized for expression in a mammalian cell. In one aspect, entire CAR construct of the disclosure is encoded by a nucleic acid molecule whose entire sequence has been codon optimized for expression in a mammalian cell. Codon optimization refers to the discovery that the frequency of occurrence of synonymous codons (i.e., codons that code for the same amino acid) in coding DNA is biased in different species. Such codon degeneracy allows an identical polypeptide to be encoded by a variety of nucleotide sequences. A variety of codon optimization methods is known in the art, and include, e.g., methods disclosed in at least U.S. Pat. Nos. 5,786,464 and 6,114,148.

Antigen Binding Domains (and the Targeted Antigens)

In one embodiment, an antigen binding domain against CD19 is an antigen binding portion, e.g., CDRs, of a CAR, antibody or antigen-binding fragment thereof described in, e.g., PCT publication WO2012/079000; PCT publication WO2014/153270; Kochenderfer, J. N. et al., J. Immunother. 32 (7), 689-702 (2009); Kochenderfer, J. N., et al., Blood, 116 (20), 4099-4102 (2010); PCT publication WO2014/031687; Bejcek, Cancer Research, 55, 2346-2351, 1995; or U.S. Pat. No. 7,446,190.

In one embodiment, an antigen binding domain against mesothelin is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment or CAR described in, e.g., PCT publication WO2015/090230. In one embodiment, an antigen binding domain against mesothelin is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment, or CAR described in, e.g., PCT publication WO1997/025068, WO1999/028471, WO2005/014652, WO2006/099141, WO2009/045957, WO2009/068204, WO2013/142034, WO2013/040557, or WO2013/063419. In one embodiment, an antigen binding domain against mesothelin is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment, or CAR described in WO/2015/090230.

In one embodiment, an antigen binding domain against CD123 is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment or CAR described in, e.g., PCT publication WO2014/130635. In one embodiment, an antigen binding domain against CD123 is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment, or CAR described in, e.g., PCT publication WO2014/138805, WO2014/138819, WO2013/173820, WO2014/144622, WO2001/66139, WO2010/126066, WO2014/144622, or US2009/0252742. In one embodiment, an antigen binding domain against CD123 is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment, or CAR described in WO/2016/028896.

In one embodiment, an antigen binding domain against EGFRvIII is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment or CAR described in, e.g., WO/2014/130657.

In one embodiment, an antigen binding domain against CD22 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Haso et al., Blood, 121(7): 1165-1174 (2013); Wayne et al., Clin Cancer Res 16(6): 1894-1903 (2010); Kato et al., Leuk Res 37(1):83-88 (2013); Creative BioMart (creativebiomart.net): MOM-18047-S(P).

In one embodiment, an antigen binding domain against CS-1 is an antigen binding portion, e.g., CDRs, of Elotuzumab (BMS), see e.g., Tai et al., 2008, Blood 112(4):1329-37; Tai et al., 2007, Blood. 110(5):1656-63.

In one embodiment, an antigen binding domain against CLL-1 is an antigen binding portion, e.g., CDRs, of an antibody available from R&D, ebiosciences, Abcam, for example, PE-CLL1-hu Cat #353604 (BioLegend); and PE-CLL1 (CLEC12A) Cat #562566 (BD). In one embodiment, an antigen binding domain against CLL-1 is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment, or CAR described in WO/2016/014535.

In one embodiment, an antigen binding domain against CD33 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Bross et al., Clin Cancer Res 7(6):1490-1496 (2001) (Gemtuzumab Ozogamicin, hP67.6), Caron et al., Cancer Res 52(24):6761-6767 (1992) (Lintuzumab, HuM195), Lapusan et al., Invest New Drugs 30(3):1121-1131 (2012) (AVE9633), Aigner et al., Leukemia 27(5): 1107-1115 (2013) (AMG330, CD33 BiTE), Dutour et al., Adv hematol 2012:683065 (2012), and Pizzitola et al., Leukemia doi:10.1038/Lue.2014.62 (2014). In one embodiment, an antigen binding domain against CD33 is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment, or CAR described in WO/2016/014576.

In one embodiment, an antigen binding domain against GD2 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Mujoo et al., Cancer Res. 47(4):1098-1104 (1987); Cheung et al., Cancer Res 45(6):2642-2649 (1985), Cheung et al., J Clin Oncol 5(9):1430-1440 (1987), Cheung et al., J Clin Oncol 16(9):3053-3060 (1998), Handgretinger et al., Cancer Immunol Immunother 35(3):199-204 (1992). In some embodiments, an antigen binding domain against GD2 is an antigen binding portion of an antibody selected from mAb 14.18, 14G2a, ch14.18, hu14.18, 3F8, hu3F8, 3G6, 8B6, 60C3, 10B8, ME36.1, and 8H9, see e.g., WO2012033885, WO2013040371, WO2013192294, WO2013061273, WO2013123061, WO2013074916, and WO201385552. In some embodiments, an antigen binding domain against GD2 is an antigen binding portion of an antibody described in US Publication No.: 20100150910 or PCT Publication No.: WO 2011160119.

In one embodiment, an antigen binding domain against BCMA is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., WO2012163805, WO200112812, and WO2003062401. In one embodiment, an antigen binding domain against BCMA is an antigen binding portion, e.g., CDRs, of an antibody, antigen-binding fragment, or CAR described in WO/2016/014565.

In one embodiment, an antigen binding domain against Tn antigen is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. No. 8,440,798, Brooks et al., PNAS 107(22):10056-10061 (2010), and Stone et al., OncoImmunology 1(6):863-873(2012).

In one embodiment, an antigen binding domain against PSMA is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Parker et al., Protein Expr Purif 89(2):136-145 (2013), US 20110268656 (J591 ScFv); Frigerio et al, European J Cancer 49(9):2223-2232 (2013) (scFvD2B); WO 2006125481 (mAbs 3/A12, 3/E7 and 3/F11) and single chain antibody fragments (scFv A5 and D7).

In one embodiment, an antigen binding domain against ROR1 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Hudecek et al., Clin Cancer Res 19(12):3153-3164 (2013); WO 2011159847; and US20130101607.

In one embodiment, an antigen binding domain against FLT3 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., WO2011076922, U.S. Pat. No. 5,777,084, EP0754230, US20090297529, and several commercial catalog antibodies (R&D, ebiosciences, Abcam).

In one embodiment, an antigen binding domain against TAG72 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Hombach et al., Gastroenterology 113(4):1163-1170 (1997); and Abcam ab691.

In one embodiment, an antigen binding domain against FAP is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Ostermann et al., Clinical Cancer Research 14:4584-4592 (2008) (FAPS), US Pat. Publication No. 2009/0304718; sibrotuzumab (see e.g., Hofheinz et al., Oncology Research and Treatment 26(1), 2003); and Tran et al., J Exp Med 210(6):1125-1135 (2013).

In one embodiment, an antigen binding domain against CD38 is an antigen binding portion, e.g., CDRs, of daratumumab (see, e.g., Groen et al., Blood 116(21):1261-1262 (2010); MOR202 (see, e.g., U.S. Pat. No. 8,263,746); or antibodies described in U.S. Pat. No. 8,362,211.

In one embodiment, an antigen binding domain against CD44v6 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Casucci et al., Blood 122(20):3461-3472 (2013).

In one embodiment, an antigen binding domain against CEA is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Chmielewski et al., Gastoenterology 143(4):1095-1107 (2012).

In one embodiment, an antigen binding domain against EPCAM is an antigen binding portion, e.g., CDRS, of an antibody selected from MT110, EpCAM-CD3 bispecific Ab (see, e.g., clinicaltrials.gov/ct2/show/NCT00635596); Edrecolomab; 3622W94; ING-1; and adecatumumab (MT201).

In one embodiment, an antigen binding domain against PRSS21 is an antigen binding portion, e.g., CDRs, of an antibody described in U.S. Pat. No. 8,080,650.

In one embodiment, an antigen binding domain against B7H3 is an antigen binding portion, e.g., CDRs, of an antibody MGA271 (Macrogenics).

In one embodiment, an antigen binding domain against KIT is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. No. 7,915,391, US20120288506, and several commercial catalog antibodies.

In one embodiment, an antigen binding domain against IL-13Ra2 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., WO2008/146911, WO2004087758, several commercial catalog antibodies, and WO2004087758.

In one embodiment, an antigen binding domain against CD30 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. No. 7,090,843 B1, and EP0805871.

In one embodiment, an antigen binding domain against GD3 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. Nos. 7,253,263; 8,207,308; US 20120276046; EP1013761; WO2005035577; and U.S. Pat. No. 6,437,098.

In one embodiment, an antigen binding domain against CD171 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Hong et al., J Immunother 37(2):93-104 (2014).

In one embodiment, an antigen binding domain against IL-11Ra is an antigen binding portion, e.g., CDRs, of an antibody available from Abcam (cat #ab55262) or Novus Biologicals (cat #EPR5446). In another embodiment, an antigen binding domain again IL-11Ra is a peptide, see, e.g., Huang et al., Cancer Res 72(1):271-281 (2012).

In one embodiment, an antigen binding domain against PSCA is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Morgenroth et al., Prostate 67(10):1121-1131 (2007) (scFv 7F5); Nejatollahi et al., J of Oncology 2013(2013), article ID 839831 (scFv C5-II); and US Pat Publication No. 20090311181.

In one embodiment, an antigen binding domain against VEGFR2 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Chinnasamy et al., J Clin Invest 120(11):3953-3968 (2010).

In one embodiment, an antigen binding domain against LewisY is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Kelly et al., Cancer Biother Radiopharm 23(4):411-423 (2008) (hu3S193 Ab (scFvs)); Dolezal et al., Protein Engineering 16(1):47-56 (2003) (NC10 scFv).

In one embodiment, an antigen binding domain against CD24 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Maliar et al., Gastroenterology 143(5):1375-1384 (2012).

In one embodiment, an antigen binding domain against PDGFR-beta is an antigen binding portion, e.g., CDRs, of an antibody Abcam ab32570.

In one embodiment, an antigen binding domain against SSEA-4 is an antigen binding portion, e.g., CDRs, of antibody MC813 (Cell Signalling), or other commercially available antibodies.

In one embodiment, an antigen binding domain against CD20 is an antigen binding portion, e.g., CDRs, of the antibody Rituximab, Ofatumumab, Ocrelizumab, Veltuzumab, or GA101.

In one embodiment, an antigen binding domain against Folate receptor alpha is an antigen binding portion, e.g., CDRs, of the antibody IMGN853, or an antibody described in US20120009181; U.S. Pat. No. 4,851,332, LK26: U.S. Pat. No. 5,952,484.

In one embodiment, an antigen binding domain against ERBB2 (Her2/neu) is an antigen binding portion, e.g., CDRs, of the antibody trastuzumab, or pertuzumab.

In one embodiment, an antigen binding domain against MUC1 is an antigen binding portion, e.g., CDRs, of the antibody SAR566658.

In one embodiment, the antigen binding domain against EGFR is antigen binding portion, e.g., CDRs, of the antibody cetuximab, panitumumab, zalutumumab, nimotuzumab, or matuzumab.

In one embodiment, an antigen binding domain against NCAM is an antigen binding portion, e.g., CDRs, of the antibody clone 2-2B: MAB5324 (EMD Millipore).

In one embodiment, an antigen binding domain against Ephrin B2 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Abengozar et al., Blood 119(19):4565-4576 (2012).

In one embodiment, an antigen binding domain against IGF-I receptor is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. No. 8,344,112 B2; EP2322550 A1; WO 2006/138315, or PCT/US2006/022995.

In one embodiment, an antigen binding domain against CAIX is an antigen binding portion, e.g., CDRs, of the antibody clone 303123 (R&D Systems).

In one embodiment, an antigen binding domain against LMP2 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. No. 7,410,640, or US20050129701.

In one embodiment, an antigen binding domain against gp100 is an antigen binding portion, e.g., CDRs, of the antibody HMB45, NKIbetaB, or an antibody described in WO2013165940, or US20130295007

In one embodiment, an antigen binding domain against tyrosinase is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. No. 5,843,674; or US19950504048.

In one embodiment, an antigen binding domain against EphA2 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Yu et al., Mol Ther 22(1):102-111 (2014).

In one embodiment, an antigen binding domain against GD3 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. Nos. 7,253,263; 8,207,308; US 20120276046; EP1013761 A3; 20120276046; WO2005035577; or U.S. Pat. No. 6,437,098.

In one embodiment, an antigen binding domain against fucosyl GM1 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., US20100297138; or WO2007/067992.

In one embodiment, an antigen binding domain against sLe is an antigen binding portion, e.g., CDRs, of the antibody G193 (for lewis Y), see Scott A M et al, Cancer Res 60: 3254-61 (2000), also as described in Neeson et al, J Immunol May 2013 190 (Meeting Abstract Supplement) 177.10.

In one embodiment, an antigen binding domain against GM3 is an antigen binding portion, e.g., CDRs, of the antibody CA 2523449 (mAb 14F7).

In one embodiment, an antigen binding domain against HMWMAA is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Kmiecik et al., Oncoimmunology 3(1):e27185 (2014) (PMID: 24575382) (mAb9.2.27); U.S. Pat. No. 6,528,481; WO2010033866; or US 20140004124.

In one embodiment, an antigen binding domain against o-acetyl-GD2 is an antigen binding portion, e.g., CDRs, of the antibody 8B6.

In one embodiment, an antigen binding domain against TEM 1/CD248 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Marty et al., Cancer Lett 235(2):298-308 (2006); Zhao et al., J Immunol Methods 363(2):221-232 (2011).

In one embodiment, an antigen binding domain against CLDN6 is an antigen binding portion, e.g., CDRs, of the antibody IMAB027 (Ganymed Pharmaceuticals), see e.g., clinicaltrial.gov/show/NCT02054351.

In one embodiment, an antigen binding domain against TSHR is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. Nos. 8,603,466; 8,501,415; or U.S. Pat. No. 8,309,693.

In one embodiment, an antigen binding domain against GPRC5D is an antigen binding portion, e.g., CDRs, of the antibody FAB6300A (R&D Systems); or LS-A4180 (Lifespan Biosciences).

In one embodiment, an antigen binding domain against CD97 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., U.S. Pat. No. 6,846,911; de Groot et al., J Immunol 183(6):4127-4134 (2009);

or an antibody from R&D:MAB3734.

In one embodiment, an antigen binding domain against ALK is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Mino-Kenudson et al., Clin Cancer Res 16(5):1561-1571 (2010).

In one embodiment, an antigen binding domain against polysialic acid is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Nagae et al., J Biol Chem 288(47):33784-33796 (2013).

In one embodiment, an antigen binding domain against PLAC 1 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Ghods et al., Biotechnol Appl Biochem 2013 doi: 10.1002/bab.1177.

In one embodiment, an antigen binding domain against GloboH is an antigen binding portion of the antibody VK9; or an antibody described in, e.g., Kudryashov V et al, Glycoconj J. 15(3):243-9 (1998), Lou et al., Proc Natl Acad Sci USA 111(7):2482-2487 (2014); MBr1: Bremer E-G et al. J Biol Chem 259:14773-14777 (1984).

In one embodiment, an antigen binding domain against NY-BR-1 is an antigen binding portion, e.g., CDRs of an antibody described in, e.g., Jager et al., Appl Immunohistochem Mol Morphol 15(1):77-83 (2007).

In one embodiment, an antigen binding domain against WT-1 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Dao et al., Sci Transl Med 5(176):176ra33 (2013); or WO2012/135854.

In one embodiment, an antigen binding domain against MAGE-A1 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Willemsen et al., J Immunol 174(12):7853-7858 (2005) (TCR-like scFv).

In one embodiment, an antigen binding domain against sperm protein 17 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Song et al., Target Oncol 2013 Aug. 14 (PMID: 23943313); Song et al., Med Oncol 29(4):2923-2931 (2012).

In one embodiment, an antigen binding domain against Tie 2 is an antigen binding portion, e.g., CDRs, of the antibody AB33 (Cell Signalling Technology).

In one embodiment, an antigen binding domain against MAD-CT-2 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., PMID: 2450952; U.S. Pat. No. 7,635,753.

In one embodiment, an antigen binding domain against Fos-related antigen 1 is an antigen binding portion, e.g., CDRs, of the antibody 12F9 (Novus Biologicals).

In one embodiment, an antigen binding domain against MelanA/MART1 is an antigen binding portion, e.g., CDRs, of an antibody described in, EP2514766 A2; or U.S. Pat. No. 7,749,719.

In one embodiment, an antigen binding domain against sarcoma translocation breakpoints is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Luo et al, EMBO Mol. Med. 4(6):453-461 (2012).

In one embodiment, an antigen binding domain against TRP-2 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Wang et al, J Exp Med. 184(6):2207-16 (1996).

In one embodiment, an antigen binding domain against CYP1B1 is an antigen binding portion, e.g., CDRs, of an antibody described in, e.g., Maecker et al, Blood 102 (9): 3287-3294 (2003).

In one embodiment, an antigen binding domain against RAGE-1 is an antigen binding portion, e.g., CDRs, of the antibody MAB5328 (EMD Millipore).

In one embodiment, an antigen binding domain against human telomerase reverse transcriptase is an antigen binding portion, e.g., CDRs, of the antibody cat no: LS-B95-100 (Lifespan Biosciences) In one embodiment, an antigen binding domain against intestinal carboxyl esterase is an antigen binding portion, e.g., CDRs, of the antibody 4F12: cat no: LS-B6190-50 (Lifespan Biosciences).

In one embodiment, an antigen binding domain against mut hsp70-2 is an antigen binding portion, e.g., CDRs, of the antibody Lifespan Biosciences: monoclonal: cat no: LS-C133261-100 (Lifespan Biosciences).

In one embodiment, an antigen binding domain against CD79a is an antigen binding portion, e.g., CDRs, of the antibody Anti-CD79a antibody [HM47/A9] (ab3121), available from Abcam; antibody CD79A Antibody #3351 available from Cell Signalling Technology; or antibody HPA017748—Anti-CD79A antibody produced in rabbit, available from Sigma Aldrich.

In one embodiment, an antigen binding domain against CD79b is an antigen binding portion, e.g., CDRs, of the antibody polatuzumab vedotin, anti-CD79b described in Dornan et al., “Therapeutic potential of an anti-CD79b antibody-drug conjugate, anti-CD79b-vc-MMAE, for the treatment of non-Hodgkin lymphoma” Blood. 2009 Sep. 24; 114(13):2721-9. doi: 10.1182/blood-2009-02-205500. Epub 2009 Jul. 24, or the bispecific antibody Anti-CD79b/CD3 described in “4507 Pre-Clinical Characterization of T Cell-Dependent Bispecific Antibody Anti-CD79b/CD3 As a Potential Therapy for B Cell Malignancies” Abstracts of 56^(th) ASH Annual Meeting and Exposition, San Francisco, Calif. Dec. 6-9, 2014.

In one embodiment, an antigen binding domain against CD72 is an antigen binding portion, e.g., CDRs, of the antibody J3-109 described in Myers, and Uckun, “An anti-CD72 immunotoxin against therapy-refractory B-lineage acute lymphoblastic leukemia.” Leuk Lymphoma. 1995 June; 18(1-2):119-22, or anti-CD72 (10D6.8.1, mIgG1) described in Polson et al., “Antibody-Drug Conjugates for the Treatment of Non-Hodgkin's Lymphoma: Target and Linker-Drug Selection” Cancer Res Mar. 15, 2009 69; 2358.

In one embodiment, an antigen binding domain against LAIR1 is an antigen binding portion, e.g., CDRs, of the antibody ANT-301 LAIR1 antibody, available from ProSpec; or anti-human CD305 (LAIR1) Antibody, available from BioLegend.

In one embodiment, an antigen binding domain against FCAR is an antigen binding portion, e.g., CDRs, of the antibody CD89/FCARAntibody (Catalog #10414-H08H), available from Sino Biological Inc.

In one embodiment, an antigen binding domain against LILRA2 is an antigen binding portion, e.g., CDRs, of the antibody LILRA2 monoclonal antibody (M17), clone 3C₇, available from Abnova, or Mouse Anti-LILRA2 antibody, Monoclonal (2D7), available from Lifespan Biosciences.

In one embodiment, an antigen binding domain against CD300LF is an antigen binding portion, e.g., CDRs, of the antibody Mouse Anti-CMRF35-like molecule 1 antibody, Monoclonal[UP-D2], available from BioLegend, or Rat Anti-CMRF35-like molecule 1 antibody, Monoclonal[234903], available from R&D Systems.

In one embodiment, an antigen binding domain against CLEC12A is an antigen binding portion, e.g., CDRs, of the antibody Bispecific T cell Engager (BiTE) scFv-antibody and ADC described in Noordhuis et al., “Targeting of CLEC12A In Acute Myeloid Leukemia by Antibody-Drug-Conjugates and Bispecific CLL-1xCD3 BiTE Antibody” 53^(rd) ASH Annual Meeting and Exposition, Dec. 10-13, 2011, and MCLA-117 (Merus).

In one embodiment, an antigen binding domain against BST2 (also called CD317) is an antigen binding portion, e.g., CDRs, of the antibody Mouse Anti-CD317 antibody, Monoclonal[3H4], available from Antibodies-Online or Mouse Anti-CD317 antibody, Monoclonal[696739], available from R&D Systems.

In one embodiment, an antigen binding domain against EMR2 (also called CD312) is an antigen binding portion, e.g., CDRs, of the antibody Mouse Anti-CD312 antibody, Monoclonal[LS-B8033]available from Lifespan Biosciences, or Mouse Anti-CD312 antibody, Monoclonal[494025] available from R&D Systems.

In one embodiment, an antigen binding domain against LY75 is an antigen binding portion, e.g., CDRs, of the antibody Mouse Anti-Lymphocyte antigen 75 antibody, Monoclonal[HD30] available from EMD Millipore or Mouse Anti-Lymphocyte antigen 75 antibody, Monoclonal[A15797] available from Life Technologies.

In one embodiment, an antigen binding domain against GPC3 is an antigen binding portion, e.g., CDRs, of the antibody hGC33 described in Nakano K, Ishiguro T, Konishi H, et al. Generation of a humanized anti-glypican 3 antibody by CDR grafting and stability optimization. Anticancer Drugs. 2010 November; 21(10):907-916, or MDX-1414, HN3, or YP7, all three of which are described in Feng et al., “Glypican-3 antibodies: a new therapeutic target for liver cancer.” FEBS Lett. 2014 Jan. 21; 588(2):377-82.

In one embodiment, an antigen binding domain against FCRL5 is an antigen binding portion, e.g., CDRs, of the anti-FcRL5 antibody described in Elkins et al., “FcRL5 as a target of antibody-drug conjugates for the treatment of multiple myeloma” Mol Cancer Ther. 2012 October; 11(10):2222-32. In one embodiment, an antigen binding domain against FCRL5 is an antigen binding portion, e.g., CDRs, of the anti-FcRL5 antibody described in, for example, WO2001/038490, WO/2005/117986, WO2006/039238, WO2006/076691, WO2010/114940, WO2010/120561, or WO2014/210064.

In one embodiment, an antigen binding domain against IGLL1 is an antigen binding portion, e.g., CDRs, of the antibody Mouse Anti-Immunoglobulin lambda-like polypeptide 1 antibody, Monoclonal[AT1G4] available from Lifespan Biosciences, Mouse Anti-Immunoglobulin lambda-like polypeptide 1 antibody, Monoclonal[HSL11] available from BioLegend.

In one embodiment, the antigen binding domain comprises one, two three (e.g., all three) heavy chain CDRs, HC CDR1, HC CDR2 and HC CDR3, from an antibody listed above, and/or one, two, three (e.g., all three) light chain CDRs, LC CDR1, LC CDR2 and LC CDR3, from an antibody listed above. In one embodiment, the antigen binding domain comprises a heavy chain variable region and/or a variable light chain region of an antibody listed above.

In another aspect, the antigen binding domain comprises a humanized antibody or an antibody fragment. In some aspects, a non-human antibody is humanized, where specific sequences or regions of the antibody are modified to increase similarity to an antibody naturally produced in a human or fragment thereof. In one aspect, the antigen binding domain is humanized.

In an embodiment, the antigen-binding domain of a CAR, e.g., a CAR expressed by a cell of the disclosure, binds to CD19. CD19 is found on B cells throughout differentiation of the lineage from the pro/pre-B cell stage through the terminally differentiated plasma cell stage. In an embodiment, the antigen binding domain is a murine scFv domain that binds to human CD19, e.g., the antigen binding domain of CTL019 (e.g., SEQ ID NO: 218). In an embodiment, the antigen binding domain is a humanized antibody or antibody fragment, e.g., scFv domain, derived from the murine CTL019 scFv. In an embodiment, the antigen binding domain is a human antibody or antibody fragment that binds to human CD19. Exemplary scFv domains (and their sequences, e.g., CDRs, VL and VH sequences) that bind to CD19 are provided in Table 12a. The scFv domain sequences provided in Table 12a include a light chain variable region (VL) and a heavy chain variable region (VH). The VL and VH are attached by a linker comprising the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 216), e.g., in the following orientation: VL-linker-VH.

TABLE 12a Antigen Binding domains that bind CD19 SEQ ID Antigen Name Amino Acid Sequence NO: CD19 muCTL019 DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTV 218 KLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQ GNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEVKLQESGPGL VAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSE TTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHY YYGGSYAMDYWGQGTSVTVSS CD19 huscFv1 EIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQKPGQAP 219 RLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFAVYFCQQ GNTLPYTFGQGTKLEIKGGGGSGGGGSGGGGSQVQLQESGPGL VKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKGLEWIGVIWGSE TTYYSSSLKSRVTISKDNSKNQVSLKLSSVTAADTAVYYCAKH YYYGGSYAMDYWGQGTLVTVSS CD19 huscFv2 EIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQKPGQAP 220 RLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFAVYFCQQ GNTLPYTFGQGTKLEIKGGGGSGGGGSGGGGSQVQLQESGPGL VKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKGLEWIGVIWGSE TTYYQSSLKSRVTISKDNSKNQVSLKLSSVTAADTAVYYCAKH YYYGGSYAMDYWGQGTLVTVSS CD19 huscFv3 QVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKG 221 LEWIGVIWGSETTYYSSSLKSRVTISKDNSKNQVSLKLSSVTAA DTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSGGGGSGGGG SGGGGSEIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQ KPGQAPRLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFA VYFCQQGNTLPYTFGQGTKLEIK CD19 huscFv4 QVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKG 222 LEWIGVIWGSETTYYQSSLKSRVTISKDNSKNQVSLKLSSVTAA DTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSGGGGSGGGG SGGGGSEIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQ KPGQAPRLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFA VYFCQQGNTLPYTFGQGTKLEIK CD19 huscFv5 EIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQKPGQAP 223 RLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFAVYFCQQ GNTLPYTFGQGTKLEIKGGGGSGGGGSGGGGSGGGGSQVQLQE SGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKGLEWIGVI WGSETTYYSSSLKSRVTISKDNSKNQVSLKLSSVTAADTAVYYC AKHYYYGGSYAMDYWGQGTLVTVSS CD19 huscFv6 EIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQKPGQAP 224 RLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFAVYFCQQ GNTLPYTFGQGTKLEIKGGGGSGGGGSGGGGSGGGGSQVQLQE SGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKGLEWIGVI WGSETTYYQSSLKSRVTISKDNSKNQVSLKLSSVTAADTAVYY CAKHYYYGGSYAMDYWGQGTLVTVSS CD19 huscFv7 QVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKG 225 LEWIGVIWGSETTYYSSSLKSRVTISKDNSKNQVSLKLSSVTAA DTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSGGGGSGGGG SGGGGSGGGGSEIVMTQSPATLSLSPGERATLSCRASQDISKYL NWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSL QPEDFAVYFCQQGNTLPYTFGQGTKLEIK CD19 huscFv8 QVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKG 226 LEWIGVIWGSETTYYQSSLKSRVTISKDNSKNQVSLKLSSVTAA DTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSGGGGSGGGG SGGGGSGGGGSEIVMTQSPATLSLSPGERATLSCRASQDISKYL NWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSL QPEDFAVYFCQQGNTLPYTFGQGTKLEIK CD19 huscFv9 EIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQKPGQAP 227 RLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFAVYFCQQ GNTLPYTFGQGTKLEIKGGGGSGGGGSGGGGSGGGGSQVQLQE SGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKGLEWIGVI WGSETTYYNSSLKSRVTISKDNSKNQVSLKLSSVTAADTAVYY CAKHYYYGGSYAMDYWGQGTLVTVSS CD19 HuscFv10 QVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKG 228 LEWIGVIWGSETTYYNSSLKSRVTISKDNSKNQVSLKLSSVTAA DTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSGGGGSGGGG SGGGGSGGGGSEIVMTQSPATLSLSPGERATLSCRASQDISKYL NWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSL QPEDFAVYFCQQGNTLPYTFGQGTKLEIK CD19 HuscFv11 EIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQKPGQAP 229 RLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFAVYFCQQ GNTLPYTFGQGTKLEIKGGGGSGGGGSGGGGSQVQLQESGPGL VKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKGLEWIGVIWGSE TTYYNSSLKSRVTISKDNSKNQVSLKLSSVTAADTAVYYCAKH YYYGGSYAMDYWGQGTLVTVSS CD19 HuscFv12 QVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSWIRQPPGKG 230 LEWIGVIWGSETTYYNSSLKSRVTISKDNSKNQVSLKLSSVTAA DTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSGGGGSGGGG SGGGGSEIVMTQSPATLSLSPGERATLSCRASQDISKYLNWYQQ KPGQAPRLLIYHTSRLHSGIPARFSGSGSGTDYTLTISSLQPEDFA VYFCQQGNTLPYTFGQGTKLEIK

The sequences of the CDR sequences of the scFv domains of the CD19 antigen binding domains provided in Table 12a are shown in Table 12b for the heavy chain variable domains and in Table 12c for the light chain variable domains. “ID” stands for the respective SEQ ID NO for each CDR.

TABLE 12b Heavy Chain Variable Domain CDRs Description FW HCDR1 ID HCDR2 ID HCDR3 ID murine_CART19 GVSLPDYGVS 306 VIWGSETTYYNSALKS 307 HYYYGGSYAMDY 231 humanized_CART19 a VH4 GVSLPDYGVS 306 VIWGSETTYY

SSLKS 308 HYYYGGSYAMDY 231 humanized_CART19 b VH4 GVSLPDYGVS 306 VIWGSETTYY

SSLKS 309 HYYYGGSYAMDY 231 humanized_CART19 c VH4 GVSLPDYGVS 306 VIWGSETTYYNSSLKS 310 HYYYGGSYAMDY 231

TABLE 12c Light Chain Variable Domain CDRs Description FW LCDR1 ID LCDR2 ID LCDR3 ID murine_CART19 RASQDISKYLN 311 HTSRLHS 312 QQGNTLPYT 232 humanized_CART19 a VK3 RASQDISKYLN 311 HTSRLHS 312 QQGNTLPYT 232 humanized_CART19 b VK3 RASQDISKYLN 311 HTSRLHS 312 QQGNTLPYT 232 humanized_CART19 c VK3 RASQDISKYLN 311 HTSRLHS 312 QQGNTLPYT 232

In an embodiment, the antigen binding domain comprises an anti-CD19 antibody, or fragment thereof, e.g., a scFv. For example, the antigen binding domain comprises a variable heavy chain and a variable light chain listed in Table 12d. The linker sequence joining the variable heavy and variable light chains can be any of the linker sequences described herein, or alternatively, can be GSTSGSGKPGSGEGSTKG (SEQ ID NO: 233). The light chain variable region and heavy chain variable region of a scFv can be, e.g., in any of the following orientations: light chain variable region-linker-heavy chain variable region or heavy chain variable region-linker-light chain variable region.

TABLE 12d Additional Anti-CD19 antibody binding domains Ab Name VH Sequence VL Sequence SJ25-C1 QVQLLESGAELVRPGSSVKISCKASGYA ELVLTQSPKFMSTSVGDRVSVTCKAS FSSYWMNWVKQRPGQGLEWIGQIYPGD QNVGTNVAWYQQKPGQSPKPLIYSA GDTNYNGKFKGQATLTADKSSSTAYMQ TYRNSGVPDRFTGSGSGTDFTLTITNV LSGLTSEDSAVYSCARKTISSVVDFYFD QSKDLADYFYFCQYNRYPYTSGGGT YWGQGTTVT (SEQ ID NO: 234) KLEIKRRS (SEQ ID NO: 235) ScFv Sequence SJ25-C1 QVQLLESGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIYPGD scFv GDTNYNGKFKGQATLTADKSSSTAYMQLSGLTSEDSAVYSCARKTISSVVDFYFD YWGQGTTVTGSTSGSGKPGSGEGSTKGELVLTQSPKFMSTSVGDRVSVTCKASQN VGTNVAWYQQKPGQSPKPLIYSATYRNSGVPDRFTGSGSGTDFTLTITNVQSKDLA DYFYFCQYNRYPYTSGGGTKLEIKRRS (SEQ ID NO: 236)

In one embodiment, the CD19 binding domain comprises one or more (e.g., all three) light chain complementary determining region 1 (LC CDR1), light chain complementary determining region 2 (LC CDR2), and light chain complementary determining region 3 (LC CDR3) of a CD19 binding domain described herein, e.g., provided in Table 12a or 15, and/or one or more (e.g., all three) heavy chain complementary determining region 1 (HC CDR1), heavy chain complementary determining region 2 (HC CDR2), and heavy chain complementary determining region 3 (HC CDR3) of a CD19 binding domain described herein, e.g., provided in Table 12a or 16. In one embodiment, the CD19 binding domain comprises one, two, or all of LC CDR1, LC CDR2, and LC CDR3 of any amino acid sequences as provided in Table 12c, incorporated herein by reference; and one, two or all of HC CDR1, HC CDR2, and HC CDR3 of any amino acid sequences as provided in Table 12b.

Any known CD19 CAR, e.g., the CD19 antigen binding domain of any known CD19 CAR, in the art can be used in accordance with the instant disclosure to construct a CAR. For example, LG-740; CD19 CAR described in the U.S. Pat. Nos. 8,399,645; 7,446,190; Xu et al., Leuk Lymphoma. 2013 54(2):255-260(2012); Cruz et al., Blood 122(17):2965-2973 (2013); Brentjens et al., Blood, 118(18):4817-4828 (2011); Kochenderfer et al., Blood 116(20):4099-102 (2010); Kochenderfer et al., Blood 122 (25):4129-39(2013); and 16th Annu Meet Am Soc Gen Cell Ther (ASGCT) (May 15-18, Salt Lake City) 2013, Abst 10. In one embodiment, an antigen binding domain against CD19 is an antigen binding portion, e.g., CDRs, of a CAR, antibody or antigen-binding fragment thereof described in, e.g., PCT publication WO2012/079000; PCT publication WO2014/153270; Kochenderfer, J. N. et al., J. Immunother. 32 (7), 689-702 (2009); Kochenderfer, J. N., et al., Blood, 116 (20), 4099-4102 (2010); PCT publication WO2014/031687; Bejcek, Cancer Research, 55, 2346-2351, 1995; or U.S. Pat. No. 7,446,190.

In an embodiment, the antigen-binding domain of CAR, e.g., a CAR expressed by a cell of the disclosure, binds to BCMA. BCMA is found preferentially expressed in mature B lymphocytes. In an embodiment, the antigen binding domain is a murine scFv domain that binds to human BCMA. In an embodiment, the antigen binding domain is a humanized antibody or antibody fragment, e.g., scFv domain that binds human BCMA. In an embodiment, the antigen binding domain is a human antibody or antibody fragment that binds to human BCMA. In embodiments, exemplary BCMA CAR constructs are generated using the VH and VL sequences from PCT Publication WO2012/0163805 (the contents of which are hereby incorporated by reference in its entirety). In embodiments, additional exemplary BCMA CAR constructs are generated using the VH and VL sequences from PCT Publication WO2016/014565 (the contents of which are hereby incorporated by reference in its entirety). In embodiments, additional exemplary BCMA CAR constructs are generated using the VH and VL sequences from PCT Publication WO2014/122144 (the contents of which are hereby incorporated by reference in its entirety). In embodiments, additional exemplary BCMA CAR constructs are generated using the CAR molecules, and/or the VH and VL sequences from PCT Publication WO2016/014789 (the contents of which are hereby incorporated by reference in its entirety). In embodiments, additional exemplary BCMA CAR constructs are generated using the CAR molecules, and/or the VH and VL sequences from PCT Publication WO2014/089335 (the contents of which are hereby incorporated by reference in its entirety). In embodiments, additional exemplary BCMA CAR constructs are generated using the CAR molecules, and/or the VH and VL sequences from PCT Publication WO2014/140248 (the contents of which are hereby incorporated by reference in its entirety).

Any known BCMA CAR, e.g., the BMCA antigen binding domain of any known BCMA CAR, in the art can be used in accordance with the instant disclosure. For example, those described herein.

Exemplary CAR Molecules

In one aspect, a CAR, e.g., a CAR expressed by the cell of the disclosure, comprises a CAR molecule comprising an antigen binding domain that binds to a B cell antigen, e.g., as described herein, such as CD19 or BCMA.

In one embodiment, the CAR comprises a CAR molecule comprising a CD19 antigen binding domain (e.g., a murine, human or humanized antibody or antibody fragment that specifically binds to CD19), a transmembrane domain, and an intracellular signalling domain (e.g., an intracellular signalling domain comprising a costimulatory domain and/or a primary signalling domain).

Exemplary CAR molecules described herein are provided in Table 12e. The CAR molecules in Table 12e comprise a CD19 antigen binding domain, e.g., an amino acid sequence of any CD19 antigen binding domain provided in Table 12a.

TABLE 12e Exemplary CD19 CAR molecules SEQ ID Antigen Name Amino Acid Sequence NO: CD19 CTL019 MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCR 237 ASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSG TDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSG GGGSGGGGSEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVS WIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFL KMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSTT TPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYI WAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQ EEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELN LGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKM AEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQAL PPR CD19 CAR1 MALPVTALLLPLALLLHAARPEIVMTQSPATLSLSPGERATLSCR 238 ASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGT DYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKGGGGSGG GGSGGGGSQVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSW IRQPPGKGLEWIGVIWGSETTYYSSSLKSRVTISKDNSKNQVSLK LSSVTAADTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSTTT PAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIW APLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEE DGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLG RREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAE AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CD19 CAR 2 MALPVTALLLPLALLLHAARPEIVMTQSPATLSLSPGERATLSCR 239 ASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGT DYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKGGGGSGG GGSGGGGSQVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSW IRQPPGKGLEWIGVIWGSETTYYQSSLKSRVTISKDNSKNQVSLK LSSVTAADTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSTTT PAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIW APLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEE DGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLG RREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAE AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR CD19 CAR 3 MALPVTALLLPLALLLHAARPQVQLQESGPGLVKPSETLSLTCT 240 VSGVSLPDYGVSWIRQPPGKGLEWIGVIWGSETTYYSSSLKSRVT ISKDNSKNQVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDYW GQGTLVTVSSGGGGSGGGGSGGGGSEIVMTQSPATLSLSPGERA TLSCRASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFSG SGSGTDYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKTT TPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYI WAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQ EEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELN LGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKM AEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQAL PPR CD19 CAR 4 MALPVTALLLPLALLLHAARPQVQLQESGPGLVKPSETLSLTCT 241 VSGVSLPDYGVSWIRQPPGKGLEWIGVIWGSETTYYQSSLKSRV TISKDNSKNQVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDY WGQGTLVTVSSGGGGSGGGGSGGGGSEIVMTQSPATLSLSPGER ATLSCRASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFS GSGSGTDYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKT TTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIY IWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQ EEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELN LGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKM AEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQAL PPR CD19 CAR 5 MALPVTALLLPLALLLHAARPEIVMTQSPATLSLSPGERATLSCR 242 ASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGT DYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKGGGGSGG GGSGGGGSGGGGSQVQLQESGPGLVKPSETLSLTCTVSGVSLPD YGVSWIRQPPGKGLEWIGVIWGSETTYYSSSLKSRVTISKDNSKN QVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDYWGQGTLVT VSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFA CDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPV QTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLY NELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQK DKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHM QALPPR CD19 CAR 6 MALPVTALLLPLALLLHAARPEIVMTQSPATLSLSPGERATLSCR 243 ASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGT DYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKGGGGSGG GGSGGGGSGGGGSQVQLQESGPGLVKPSETLSLTCTVSGVSLPD YGVSWIRQPPGKGLEWIGVIWGSETTYYQSSLKSRVTISKDNSK NQVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDYWGQGTLV TVSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDF ACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRP VQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQL YNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQ KDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALH MQALPPR CD19 CAR 7 MALPVTALLLPLALLLHAARPQVQLQESGPGLVKPSETLSLTCT 244 VSGVSLPDYGVSWIRQPPGKGLEWIGVIWGSETTYYSSSLKSRVT ISKDNSKNQVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDYW GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVMTQSPATLSLS PGERATLSCRASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIP ARFSGSGSGTDYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKL EIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFA CDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPV QTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLY NELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQK DKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHM QALPPR CD19 CAR 8 MALPVTALLLPLALLLHAARPQVQLQESGPGLVKPSETLSLTCT 245 VSGVSLPDYGVSWIRQPPGKGLEWIGVIWGSETTYYQSSLKSRV TISKDNSKNQVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDY WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVMTQSPATLS LSPGERATLSCRASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSG IPARFSGSGSGTDYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGT KLEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLD FACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMR PVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQ LYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNEL QKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDAL HMQALPPR CD19 CAR 9 MALPVTALLLPLALLLHAARPEIVMTQSPATLSLSPGERATLSCR 246 ASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGT DYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKGGGGSGG GGSGGGGSGGGGSQVQLQESGPGLVKPSETLSLTCTVSGVSLPD YGVSWIRQPPGKGLEWIGVIWGSETTYYNSSLKSRVTISKDNSK NQVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDYWGQGTLV TVSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDF ACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRP VQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQL YNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQ KDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALH MQALPPR CD19 CAR 10 MALPVTALLLPLALLLHAARPEIVMTQSPATLSLSPGERATLSCR 247 ASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGT DYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKGGGGSGG GGSGGGGSGGGGSQVQLQESGPGLVKPSETLSLTCTVSGVSLPD YGVSWIRQPPGKGLEWIGVIWGSETTYYNSSLKSRVTISKDNSK NQVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDYWGQGTLV TVSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDF ACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRP VQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQL YNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQ KDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALH MQALPPR CD19 CAR 11 MALPVTALLLPLALLLHAARPQVQLQESGPGLVKPSETLSLTCT 248 VSGVSLPDYGVSWIRQPPGKGLEWIGVIWGSETTYYNSSLKSRV TISKDNSKNQVSLKLSSVTAADTAVYYCAKHYYYGGSYAMDY WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVMTQSPATLS LSPGERATLSCRASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSG IPARFSGSGSGTDYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGT KLEIKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLD FACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMR PVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQ LYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNEL QKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDAL HMQALPPR CD19 CAR 12 MALPVTALLLPLALLLHAARPEIVMTQSPATLSLSPGERATLSCR 249 ASQDISKYLNWYQQKPGQAPRLLIYHTSRLHSGIPARFSGSGSGT DYTLTISSLQPEDFAVYFCQQGNTLPYTFGQGTKLEIKGGGGSGG GGSGGGGSQVQLQESGPGLVKPSETLSLTCTVSGVSLPDYGVSW IRQPPGKGLEWIGVIWGSETTYYNSSLKSRVTISKDNSKNQVSLK LSSVTAADTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSTTT PAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIW APLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEE DGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLG RREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAE AYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

In one aspect, a CAR, e.g., a CAR expressed by the cell of the disclosure, comprises a CAR molecule comprising an antigen binding domain that binds to BCMA, e.g., comprises a BCMA antigen binding domain (e.g., a murine, human or humanized antibody or antibody fragment that specifically binds to BCMA, e.g., human BCMA), a transmembrane domain, and an intracellular signalling domain (e.g., an intracellular signalling domain comprising a costimulatory domain and/or a primary signalling domain).

Exemplary CAR molecules of a CAR described herein are provided in Table 1 of WO2016/014565, which is incorporated by reference herein.

Transmembrane Domains

With respect to the transmembrane domain, in various embodiments, a CAR can be designed to comprise a transmembrane domain that is attached to the extracellular domain of the CAR. A transmembrane domain can include one or more additional amino acids adjacent to the transmembrane region, e.g., one or more amino acid associated with the extracellular region of the protein from which the transmembrane was derived (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 up to 15 amino acids of the extracellular region) and/or one or more additional amino acids associated with the intracellular region of the protein from which the transmembrane protein is derived (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 up to 15 amino acids of the intracellular region). In one aspect, the transmembrane domain is one that is associated with one of the other domains of the CAR e.g., in one embodiment, the transmembrane domain may be from the same protein that the signalling domain, costimulatory domain or the hinge domain is derived from. In another aspect, the transmembrane domain is not derived from the same protein that any other domain of the CAR is derived from. In some instances, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domains to the transmembrane domains of the same or different surface membrane proteins, e.g., to minimize interactions with other members of the receptor complex. In one aspect, the transmembrane domain is capable of homodimerization with another CAR on the cell surface of a CAR-expressing cell. In a different aspect, the amino acid sequence of the transmembrane domain may be modified or substituted so as to minimize interactions with the binding domains of the native binding partner present in the same CAR-expressing cell.

The transmembrane domain may be derived either from a natural or from a recombinant source. Where the source is natural, the domain may be derived from any membrane-bound or transmembrane protein. In one aspect, the transmembrane domain is capable of signalling to the intracellular domain(s) whenever the CAR has bound to a target. A transmembrane domain of particular use in this disclosure may include at least the transmembrane region(s) of e.g., the alpha, beta or zeta chain of the T-cell receptor, CD28, CD27, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154. In some embodiments, a transmembrane domain may include at least the transmembrane region(s) of, e.g., KIRDS2, OX40, CD2, CD27, LFA-1 (CD11a, CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD40, BAFFR, HVEM (LIGHTR), SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD160, CD19, IL2R beta, IL2R gamma, IL7R α, ITGA1, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, TNFR2, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, PAG/Cbp, NKG2D, NKG2C.

In some instances, the transmembrane domain can be attached to the extracellular region of the CAR, e.g., the antigen binding domain of the CAR, via a hinge, e.g., a hinge from a human protein. For example, in one embodiment, the hinge can be a human Ig (immunoglobulin) hinge (e.g., an IgG4 hinge, an IgD hinge), a GS linker (e.g., a GS linker described herein), a KIR2DS2 hinge or a CD8a hinge. In one embodiment, the hinge or spacer comprises (e.g., consists of) the amino acid sequence of SEQ ID NO: 250. In one aspect, the transmembrane domain comprises (e.g., consists of) a transmembrane domain of SEQ ID NO: 251.

In certain embodiments, the encoded transmembrane domain comprises an amino acid sequence of a CD8 transmembrane domain having at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO: 251, or a sequence with at least 95% identity to the amino acid sequence of SEQ ID NO: 251. In one embodiment, the encoded transmembrane domain comprises the sequence of SEQ ID NO: 251.

In other embodiments, the nucleic acid molecule encoding the CAR comprises a nucleotide sequence of a CD8 transmembrane domain, e.g., comprising the sequence of SEQ ID NO: 252 or SEQ ID NO: 289, or a sequence with at least 95% identity thereof.

In certain embodiments, the encoded antigen binding domain is connected to the transmembrane domain by a hinge region. In one embodiment, the encoded hinge region comprises the amino acid sequence of a CD8 hinge, e.g., SEQ ID NO: 250; or the amino acid sequence of an IgG4 hinge, e.g., SEQ ID NO: 253 or a sequence with at least 95% identity to SEQ ID NO: 250 or SEQ ID NO: 253. In other embodiments, the nucleic acid sequence encoding the hinge region comprises the sequence of SEQ ID NO: 254 or SEQ ID NO: 255, corresponding to a CD8 hinge or an IgG4 hinge, respectively, or a sequence with at least 95% identity to SEQ ID NO: 254 or 255.

In one aspect, the hinge or spacer comprises an IgG4 hinge. For example, in one embodiment, the hinge or spacer comprises a hinge of the amino acid sequence ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQ VYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKM (SEQ ID NO: 253). In some embodiments, the hinge or spacer comprises a hinge encoded by the nucleotide sequence of GAGAGCAAGTACGGCCCTCCCTGCCCCCCTTGCCCTGCCCCCGAGTTCCTGGGCGGACCCAG CGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAGGTGA CCTGTGTGGTGGTGGACGTGTCCCAGGAGGACCCCGAGGTCCAGTTCAACTGGTACGTGGAC GGCGTGGAGGTGCACAACGCCAAGACCAAGCCCCGGGAGGAGCAGTTCAATAGCACCTACC GGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAATACAAGTG TAAGGTGTCCAACAAGGGCCTGCCCAGCAGCATCGAGAAAACCATCAGCAAGGCCAAGGGC CAGCCTCGGGAGCCCCAGGTGTACACCCTG CCCTAGCCAAGAGGAGATGACCAAGAACC AGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAG AGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCA GCTTCTTCCTGTACAGCCGGCTGACCGTGGACAAGAGCCGGTGGCAGGAGGGCAACGTCTTT AGCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTC CCTGGGCAAGATG (SEQ ID NO: 255).

In one aspect, the hinge or spacer comprises an IgD hinge. For example, in one embodiment, the hinge or spacer comprises a hinge of the amino acid sequence of RWPESPKAQASSVPTAQPQAEGSLAKATTAPATTRNTGRGGEEKKKEKEKEEQEERETKTPECP SHTQPLGVYLLTPAVQDLWLRDKATFTCFVVGSDLKDAHLTWEVAGKVPTGGVEEGLLERHSN GSQSQHSRLTLPRSLWNAGTSVTCTLNHPSLPPQRLMALREPAAQAPVKLSLNLLASSDPPEAAS WLLCEVSGFSPPNILLMWLEDQREVNTSGFAPARPPPQPGSTTFWAWSVLRVPAPPSPQPATYTC VVSHEDSRTLLNASRSLEVSYVTDH (SEQ ID NO: 256). In some embodiments, the hinge or spacer comprises a hinge encoded by the nucleotide sequence of AGGTGGCCCGAAAGTCCCAAGGCCCAGGCATCTAGTGTTCCTACTGCACAGCCCCAGGCAG AAGGCAGCCTAGCCAAAGCTACTACTGCACCTGCCACTACGCGCAATACTGGCCGTGGCGG GGAGGAGAAGAAAAAGGAGAAAGAGAAAGAAGAACAGGAAGAGAGGGAGACCAAGACCC CTGAATGTCCATCCCATACCCAGCCGCTGGGCGTCTATCTCTTGACTCCCGCAGTACAGGAC TTGTGGCTTAGAGATAAGGCCACCTTTACATGTTTCGTCGTGGGCTCTGACCTGAAGGATGC CCATTTGACTTGGGAGGTTGCCGGAAAGGTACCCACAGGGGGGGTTGAGGAAGGGTTGCTG GAGCGCCATTCCAATGGCTCTCAGAGCCAGCACTCAAGACTCACCCTTCCGAGATCCCTGTG GAACGCCGGGACCTCTGTCACATGTACTCTAAATCATCCTAGCCTGCCCCCACAGCGTCTGA TGGCCCTTAGAGAGCCAGCCGCCCAGGCACCAGTTAAGCTTAGCCTGAATCTGCTCGCCAGT AGTGATCCCCCAGAGGCCGCCAGCTGGCTCTTATGCGAAGTGTCCGGCTTTAGCCCGCCCAA CATCTTGCTCATGTGGCTGGAGGACCAGCGAGAAGTGAACACCAGCGGCTTCGCTCCAGCCC GGCCCCCACCCCAGCCGGGTTCTACCACATTCTGGGCCTGGAGTGTCTTAAGGGTCCCAGCA CCACCTAGCCCCCAGCCAGCCACATACACCTGTGTTGTGTCCCATGAAGATAGCAGGACCCT GCTAAATGCTTCTAGGAGTCTGGAGGTTTCCTACGTGACTGACCATT (SEQ ID NO: 257).

In one aspect, the transmembrane domain may be recombinant, in which case it will comprise predominantly hydrophobic residues such as leucine and valine. In one aspect a triplet of phenylalanine, tryptophan and valine can be found at each end of a recombinant transmembrane domain.

Optionally, a short oligo- or polypeptide linker, between 2 and 10 amino acids in length may form the linkage between the transmembrane domain and the cytoplasmic region of the CAR. A glycine-serine doublet provides a particularly suitable linker. For example, in one aspect, the linker comprises the amino acid sequence of GGGGSGGGGS (SEQ ID NO: 258). In some embodiments, the linker is encoded by the nucleotide sequence of GGTGGCGGAGGTTCTGGAGGTGGAGGTTCC (SEQ ID NO: 259).

In one aspect, the hinge or spacer comprises a KIR2DS2 hinge.

Signalling Domains

In embodiments of the disclosure having an intracellular signalling domain, such a domain can contain, e.g., one or more of a primary signalling domain and/or a costimulatory signalling domain. In some embodiments, the intracellular signalling domain comprises a sequence encoding a primary signalling domain. In some embodiments, the intracellular signalling domain comprises a costimulatory signalling domain. In some embodiments, the intracellular signalling domain comprises a primary signalling domain and a costimulatory signalling domain.

The intracellular signalling sequences within the cytoplasmic portion of the CAR of the disclosure may be linked to each other in a random or specified order. Optionally, a short oligo- or polypeptide linker, for example, between 2 and 10 amino acids (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids) in length may form the linkage between intracellular signalling sequences. In one embodiment, a glycine-serine doublet can be used as a suitable linker. In one embodiment, a single amino acid, e.g., an alanine, a glycine, can be used as a suitable linker.

In one aspect, the intracellular signalling domain is designed to comprise two or more, e.g., 2, 3, 4, 5, or more, costimulatory signalling domains. In an embodiment, the two or more, e.g., 2, 3, 4, 5, or more, costimulatory signalling domains, are separated by a linker molecule, e.g., a linker molecule described herein. In one embodiment, the intracellular signalling domain comprises two costimulatory signalling domains. In some embodiments, the linker molecule is a glycine residue. In some embodiments, the linker is an alanine residue.

Primary Signalling Domains

A primary signalling domain regulates primary activation of the TCR complex either in a stimulatory way, or in an inhibitory way. Primary intracellular signalling domains that act in a stimulatory manner may contain signalling motifs, which are known as immunoreceptor tyrosine-based activation motifs or ITAMs.

Examples of ITAM containing primary intracellular signalling domains that are of particular use in the disclosure include those of CD3 zeta, common FcR gamma (FCER1G), Fc gamma RIIa, FcR beta (Fc Epsilon R1b), CD3 gamma, CD3 delta, CD3 epsilon, CD79a, CD79b, DAP10, and DAP12. In one embodiment, a CAR of the disclosure comprises an intracellular signalling domain, e.g., a primary signalling domain of CD3-zeta.

In one embodiment, the encoded primary signalling domain comprises a functional signalling domain of CD3 zeta. The encoded CD3 zeta primary signalling domain can comprise an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO: 260 or SEQ ID NO: 261, or a sequence with at least 95% identity to the amino acid sequence of SEQ ID NO: 260 or SEQ ID NO: 261. In some embodiments, the encoded primary signalling domain comprises the sequence of SEQ ID NO: 260 or SEQ ID NO: 261. In other embodiments, the nucleic acid sequence encoding the primary signalling domain comprises the sequence of SEQ ID NO: 262, SEQ ID NO: 291, or SEQ ID NO: 263, or a sequence with at least 95% identity thereof.

Costimulatory Signalling Domains

In some embodiments, the encoded intracellular signalling domain comprises a costimulatory signalling domain. For example, the intracellular signalling domain can comprise a primary signalling domain and a costimulatory signalling domain. In some embodiments, the encoded costimulatory signalling domain comprises a functional signalling domain of a protein chosen from one or more of CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, CDS, ICAM-1, GITR, BAFFR, HVEM (LIGHTR), SLAMF7, NKp80 (KLRF1), CD160, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, NKp44, NKp30, NKp46, or NKG2D.

In certain embodiments, the encoded costimulatory signalling domain comprises an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO: 264 or SEQ ID NO: 265, or a sequence with at least 95% identity to the amino acid sequence of SEQ ID NO: 264 or SEQ ID NO: 265. In one embodiment, the encoded costimulatory signalling domain comprises the sequence of SEQ ID NO: 264 or SEQ ID NO: 265. In other embodiments, the nucleic acid sequence encoding the costimulatory signalling domain comprises the sequence of SEQ ID NO: 266, SEQ ID NO: 290, or SEQ ID NO: 267, or a sequence with at least 95% identity thereof.

In other embodiments, the encoded intracellular domain comprises the sequence of SEQ ID NO: 264 or SEQ ID NO: 265 and the sequence of SEQ ID NO: 260 or SEQ ID NO: 261, wherein the sequences comprising the intracellular signalling domain are expressed in the same frame and as a single polypeptide chain.

In certain embodiments, the nucleic acid sequence encoding the intracellular signalling domain comprises the sequence of SEQ ID NO: 266, SEQ ID NO: 290, or SEQ ID NO: 267, or a sequence with at least 95% identity thereof, and the sequence of SEQ ID NO: 262, SEQ ID NO: 291, or SEQ ID NO: 263, or a sequence with at least 95% identity thereof.

In some embodiments, the nucleic acid molecule further encodes a leader sequence. In one embodiment, the leader sequence comprises the sequence of SEQ ID NO: 268.

In one aspect, the intracellular signalling domain is designed to comprise the signalling domain of CD3-zeta and the signalling domain of CD28. In one aspect, the intracellular signalling domain is designed to comprise the signalling domain of CD3-zeta and the signalling domain of 4-1BB. In one aspect, the signalling domain of 4-1BB is a signalling domain of SEQ ID NO: 264. In one aspect, the signalling domain of CD3-zeta is a signalling domain of SEQ ID NO: 260.

In one aspect, the intracellular signalling domain is designed to comprise the signalling domain of CD3-zeta and the signalling domain of CD27. In one aspect, the signalling domain of CD27 comprises the amino acid sequence of QRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP (SEQ ID NO: 265). In one aspect, the signalling domain of CD27 is encoded by the nucleic acid sequence of Caacgaaggaaatatagatcaaacaaaggagaaagtcctgtggagcctgcagagccttgtcgttacagctgccccagggaggaggagggcagcacc atccccatccaggaggattaccgaaaaccggagcctgcctgctccccc (SEQ ID NO: 267).

Vectors

In another aspect, the disclosure pertains to a vector comprising a nucleic acid sequence encoding a CAR described herein. In one embodiment, the vector is chosen from a DNA vector, an RNA vector, a plasmid, a lentivirus vector, adenoviral vector, or a retrovirus vector. In one embodiment, the vector is a lentivirus vector. These vectors or portions thereof may, among other things, be used to create template nucleic acids, as described herein for use with the CRISPR systems as described herein. Alternatively, the vectors may be used to deliver nucleic acid directly to the cell, e.g., the immune effector cell, e.g., the T cell, e.g., the allogeneic T cell, independent of the CRISPR system.

The present disclosure also provides vectors in which a DNA of the present disclosure is inserted. Vectors derived from retroviruses such as the lentivirus are suitable tools to achieve long-term gene transfer since they allow long-term, stable integration of a transgene and its propagation in daughter cells. Lentiviral vectors have the added advantage over vectors derived from onco-retroviruses such as murine leukemia viruses in that they can transduce non-proliferating cells, such as hepatocytes. They also have the added advantage of low immunogenicity. A retroviral vector may also be, e.g., a gammaretroviral vector. A gammaretroviral vector may include, e.g., a promoter, a packaging signal (ψ), a primer binding site (PBS), one or more (e.g., two) long terminal repeats (LTR), and a transgene of interest, e.g., a gene encoding a CAR. A gammaretroviral vector may lack viral structural gens such as gag, pol, and env. Exemplary gammaretroviral vectors include Murine Leukemia Virus (MLV), Spleen-Focus Forming Virus (SFFV), and Myeloproliferative Sarcoma Virus (MPSV), and vectors derived therefrom. Other gammaretroviral vectors are described, e.g., in Tobias Maetzig et al., “Gammaretroviral Vectors: Biology, Technology and Application” Viruses. 2011 June; 3(6): 677-713.

In another embodiment, the vector comprising the nucleic acid encoding the desired CAR of the disclosure is an adenoviral vector (A5/35). In another embodiment, the expression of nucleic acids encoding CARs can be accomplished using of transposons such as sleeping beauty, crisper, CAS9, and zinc finger nucleases. See below June et al. 2009 Nature Reviews Immunology 9.10: 704-716, is incorporated herein by reference.

The nucleic acid can be cloned into a number of types of vectors. For example, the nucleic acid can be cloned into a vector including, but not limited to a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors.

Disclosed herein are methods for producing an in vitro transcribed RNA CAR. The present disclosure also includes a CAR encoding RNA construct that can be directly transfected into a cell. A method for generating mRNA for use in transfection can involve in vitro transcription (IVT) of a template with specially designed primers, followed by polyA addition, to produce a construct containing 3′ and 5′ untranslated sequence (“UTR”), a 5′ cap and/or Internal Ribosome Entry Site (IRES), the nucleic acid to be expressed, and a polyA tail, typically 50-2000 bases in length (SEQ ID NO: 269). RNA so produced can efficiently transfect different kinds of cells. In one aspect, the template includes sequences for the CAR.

Non-Viral Delivery Methods

In some aspects, non-viral methods can be used to deliver a nucleic acid encoding a CAR described herein into a cell or tissue or a subject.

In some embodiments, the non-viral method includes the use of a transposon (also called a transposable element). In some embodiments, a transposon is a piece of DNA that can insert itself at a location in a genome, for example, a piece of DNA that is capable of self-replicating and inserting its copy into a genome, or a piece of DNA that can be spliced out of a longer nucleic acid and inserted into another place in a genome. For example, a transposon comprises a DNA sequence made up of inverted repeats flanking genes for transposition.

In some embodiments, cells, e.g., T or NK cells, are generated that express a CAR described herein by using a combination of gene insertion using the SBTS and genetic editing using a nuclease (e.g., Zinc finger nucleases (ZFNs), Transcription Activator-Like Effector Nucleases (TALENs), the CRISPR/Cas system, or engineered meganuclease re-engineered homing endonucleases).

In some embodiments, cells of the disclosure, e.g., T or NK cells, e.g., allogeneic T cells, e.g., described herein, (e.g., that express a CAR described herein) are generated by contacting the cells with (a) a composition comprising one or more gRNA molecules, e.g., as described herein, and one or more Cas molecules, e.g., a Cas9 molecule, e.g., as described herein, and (b) nucleic acid comprising sequence encoding a CAR, e.g., described herein (such as a template nucleic acid molecule as described herein). Without being bound by theory, said composition of (a), above, will induce a break at or near the genomic DNA targeted by the targeting domain of the gRNA molecule(s), and the nucleic acid of (b) will incorporate, e.g., partially or wholly, into the genome at or near said break, such that upon integration, the encoded CAR molecule is expressed. In embodiments, expression of the CAR will be controlled by promoters or other regulatory elements endogenous to the genome (e.g., the promoter controlling expression from the gene in which the nucleic acid of (b) was inserted). In other embodiments, the nucleic acid of (b) further comprises a promoter and/or other regulatory elements, e.g., as described herein, e.g., an EF1-alpha promoter, operably linked to the sequence encoding the CAR, such that upon integration, expression of the CAR is controlled by that promoter and/or other regulatory elements. Additional features of the disclosure relating to use of CRISPR/Cas9 systems, e.g., as described herein, to direct incorporation of nucleic acid sequence encoding a CAR, e.g., as described herein, are described elsewhere in this application, e.g., in the section relating to gene insertion and homologous recombination. In embodiments, the composition of a) above is a composition comprising RNPs comprising the one or more gRNA molecules. In embodiments, RNPs comprising gRNAs targeting unique target sequences are introduced into the cell simultaneously, e.g., as a mixture of RNPs comprising the one or more gRNAs. In embodiments, RNPs comprising gRNAs targeting unique target sequences are introduced into the cell sequentially.

In some embodiments, use of a non-viral method of delivery permits reprogramming of cells, e.g., T or NK cells, and direct infusion of the cells into a subject. Advantages of non-viral vectors include but are not limited to the ease and relatively low cost of producing sufficient amounts required to meet a patient population, stability during storage, and lack of immunogenicity.

Promoters

In one embodiment, the vector further comprises a promoter. In some embodiments, the promoter is chosen from an EF-1 promoter, a CMV IE gene promoter, an EF-1α promoter, an ubiquitin C promoter, or a phosphoglycerate kinase (PGK) promoter. In one embodiment, the promoter is an EF-1 promoter. In one embodiment, the EF-1 promoter comprises the sequence of SEQ ID NO: 270.

Host Cells for CAR Expression

As noted above, in some aspects the disclosure pertains to a cell, e.g., an immune effector cell, (e.g., a population of cells, e.g., a population of immune effector cells) comprising a nucleic acid molecule, a CAR polypeptide molecule, or a vector as described herein.

In certain aspects of the present disclosure, immune effector cells, e.g., T cells, can be obtained from a unit of blood collected from a subject using any number of techniques known to the skilled artisan, such as Ficoll™ separation. In one preferred aspect, cells from the circulating blood of an individual are obtained by apheresis. The apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets. In one aspect, the cells collected by apheresis may be washed to remove the plasma fraction and, optionally, to place the cells in an appropriate buffer or media for subsequent processing steps. In one embodiment, the cells are washed with phosphate buffered saline (PBS). In an alternative embodiment, the wash solution lacks calcium and may lack magnesium or may lack many if not all divalent cations.

Initial activation steps in the absence of calcium can lead to magnified activation. As those of ordinary skill in the art would readily appreciate a washing step may be accomplished by methods known to those in the art, such as by using a semi-automated “flow-through” centrifuge (for example, the Cobe 2991 cell processor, the Baxter CytoMate, or the Haemonetics Cell Saver 5) according to the manufacturer's instructions. After washing, the cells may be resuspended in a variety of biocompatible buffers, such as, for example, Ca-free, Mg-free PBS, PlasmaLyte A, or other saline solution with or without buffer. Alternatively, the undesirable components of the apheresis sample may be removed and the cells directly resuspended in culture media.

It is recognized that the methods of the application can utilize culture media conditions comprising 5% or less, for example 2%, human AB serum, and employ known culture media conditions and compositions, for example those described in Smith et al., “Ex vivo expansion of human T cells for adoptive immunotherapy using the novel Xeno-free CTS Immune Cell Serum Replacement” Clinical & Translational Immunology (2015) 4, e31; doi:10.1038/cti.2014.31.

In one aspect, T cells are isolated from peripheral blood lymphocytes by lysing the red blood cells and depleting the monocytes, for example, by centrifugation through a PERCOLL™ gradient or by counterflow centrifugal elutriation.

The methods described herein can include, e.g., selection of a specific subpopulation of immune effector cells, e.g., T cells, that are a T regulatory cell-depleted population, CD25+ depleted cells, using, e.g., a negative selection technique, e.g., described herein. Preferably, the population of T regulatory depleted cells contains less than 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1% of CD25+ cells.

In one embodiment, T regulatory cells, e.g., CD25+ T cells, are removed from the population using an anti-CD25 antibody, or fragment thereof, or a CD25-binding ligand, IL-2. In one embodiment, the anti-CD25 antibody, or fragment thereof, or CD25-binding ligand is conjugated to a substrate, e.g., a bead, or is otherwise coated on a substrate, e.g., a bead. In one embodiment, the anti-CD25 antibody, or fragment thereof, is conjugated to a substrate as described herein.

In one embodiment, the T regulatory cells, e.g., CD25+ T cells, are removed from the population using CD25 depletion reagent from Miltenyi™. In one embodiment, the ratio of cells to CD25 depletion reagent is 1e7 cells to 20 uL, or 1e7 cells to 15 uL, or 1e7 cells to 10 uL, or 1e7 cells to 5 uL, or 1e7 cells to 2.5 uL, or 1e7 cells to 1.25 uL. In one embodiment, e.g., for T regulatory cells, e.g., CD25+ depletion, greater than 500 million cells/ml is used. In a further aspect, a concentration of cells of 600, 700, 800, or 900 million cells/ml is used.

In one embodiment, the population of immune effector cells to be depleted includes about 6×10⁹ CD25+ T cells. In other aspects, the population of immune effector cells to be depleted include about 1×10⁹ to 1×10¹⁰ CD25+ T cell, and any integer value in between. In one embodiment, the resulting population T regulatory depleted cells has 2×10⁹ T regulatory cells, e.g., CD25+ cells, or less (e.g., 1×10⁹, 5×10⁸, 1×10⁸, 5×10⁷, 1×10⁷, or less CD25+ cells).

In one embodiment, the T regulatory cells, e.g., CD25+ cells, are removed from the population using the CliniMAC system with a depletion tubing set, such as, e.g., tubing 162-01. In one embodiment, the CliniMAC system is run on a depletion setting such as, e.g., DEPLETION2.1.

Without wishing to be bound by a particular theory, decreasing the level of negative regulators of immune cells (e.g., decreasing the number of unwanted immune cells, e.g., T_(REG) cells), in a subject prior to apheresis or during manufacturing of a CAR-expressing cell product can reduce the risk of subject relapse. For example, methods of depleting T_(REG) cells are known in the art. Methods of decreasing T_(REG) cells include, but are not limited to, cyclophosphamide, anti-GITR antibody (an anti-GITR antibody described herein), CD25-depletion, and combinations thereof.

In some embodiments, the manufacturing methods comprise reducing the number of (e.g., depleting) T_(REG) cells prior to manufacturing of the CAR-expressing cell. For example, manufacturing methods comprise contacting the sample, e.g., the apheresis sample, with an anti-GITR antibody and/or an anti-CD25 antibody (or fragment thereof, or a CD25-binding ligand), e.g., to deplete T_(REG) cells prior to manufacturing of the CAR-expressing cell (e.g., T cell, NK cell) product.

In an embodiment, a subject is pre-treated with one or more therapies that reduce T_(REG) cells prior to collection of cells for CAR-expressing cell product manufacturing, thereby reducing the risk of subject relapse to CAR-expressing cell treatment. In an embodiment, methods of decreasing T_(REG) cells include, but are not limited to, administration to the subject of one or more of cyclophosphamide, anti-GITR antibody, CD25-depletion, or a combination thereof. Administration of one or more of cyclophosphamide, anti-GITR antibody, CD25-depletion, or a combination thereof, can occur before, during or after an infusion of the CAR-expressing cell product.

In an embodiment, a subject is pre-treated with cyclophosphamide prior to collection of cells for CAR-expressing cell product manufacturing, thereby reducing the risk of subject relapse to CAR-expressing cell treatment. In an embodiment, a subject is pre-treated with an anti-GITR antibody prior to collection of cells for CAR-expressing cell product manufacturing, thereby reducing the risk of subject relapse to CAR-expressing cell treatment.

In one embodiment, the population of cells to be removed are neither the regulatory T cells or tumor cells, but cells that otherwise negatively affect the expansion and/or function of CART cells, e.g. cells expressing CD14, CD11b, CD33, CD15, or other markers expressed by potentially immune suppressive cells. In one embodiment, such cells are envisioned to be removed concurrently with regulatory T cells and/or tumor cells, or following said depletion, or in another order.

The methods described herein can include more than one selection step, e.g., more than one depletion step. Enrichment of a T cell population by negative selection can be accomplished, e.g., with a combination of antibodies directed to surface markers unique to the negatively selected cells. One method is cell sorting and/or selection via negative magnetic immunoadherence or flow cytometry that uses a cocktail of monoclonal antibodies directed to cell surface markers present on the cells negatively selected. For example, to enrich for CD4+ cells by negative selection, a monoclonal antibody cocktail can include antibodies to CD14, CD20, CD1 b, CD16, HLA-DR, and CD8.

The methods described herein can further include removing cells from the population which express a tumor antigen, e.g., a tumor antigen that does not comprise CD25, e.g., CD19, CD30, CD38, CD123, CD20, CD14 or CD11b, to thereby provide a population of T regulatory depleted, e.g., CD25+ depleted, and tumor antigen depleted cells that are suitable for expression of a CAR, e.g., a CAR described herein. In one embodiment, tumor antigen expressing cells are removed simultaneously with the T regulatory, e.g., CD25+ cells. For example, an anti-CD25 antibody, or fragment thereof, and an anti-tumor antigen antibody, or fragment thereof, can be attached to the same substrate, e.g., bead, which can be used to remove the cells or an anti-CD25 antibody, or fragment thereof, or the anti-tumor antigen antibody, or fragment thereof, can be attached to separate beads, a mixture of which can be used to remove the cells. In other embodiments, the removal of T regulatory cells, e.g., CD25+ cells, and the removal of the tumor antigen expressing cells is sequential, and can occur, e.g., in either order.

Also provided are methods that include removing cells from the population which express a check point inhibitor, e.g., a check point inhibitor described herein, e.g., one or more of PD1+ cells, LAG3+ cells, and TIM3+ cells, to thereby provide a population of T regulatory depleted, e.g., CD25+ depleted cells, and check point inhibitor depleted cells, e.g., PD1+, LAG3+ and/or TIM3+ depleted cells. Exemplary check point inhibitors include B7-H1, B7-1, CD160, P1H, 2B4, PD1, TIM3, CEACAM (e.g., CEACAM-1, CEACAM-3 and/or CEACAM-5), LAG3, TIGIT, CTLA-4, BTLA and LAIR1. In one embodiment, check point inhibitor expressing cells are removed simultaneously with the T regulatory, e.g., CD25+ cells. For example, an anti-CD25 antibody, or fragment thereof, and an anti-check point inhibitor antibody, or fragment thereof, can be attached to the same bead which can be used to remove the cells, or an anti-CD25 antibody, or fragment thereof, and the anti-check point inhibitor antibody, or fragment there, can be attached to separate beads, a mixture of which can be used to remove the cells. In other embodiments, the removal of T regulatory cells, e.g., CD25+ cells, and the removal of the check point inhibitor expressing cells is sequential, and can occur, e.g., in either order.

Methods described herein can include a positive selection step. For example, T cells can isolated by incubation with anti-CD3/anti-CD28 (e.g., 3×28)-conjugated beads, such as DYNABEADS® M-450 CD3/CD28 T, for a time period sufficient for positive selection of the desired T cells. In one embodiment, the time period is about 30 minutes. In a further embodiment, the time period ranges from 30 minutes to 36 hours or longer and all integer values there between. In a further embodiment, the time period is at least 1, 2, 3, 4, 5, or 6 hours. In yet another embodiment, the time period is 10 to 24 hours, e.g., 24 hours. Longer incubation times may be used to isolate T cells in any situation where there are few T cells as compared to other cell types, such in isolating tumor infiltrating lymphocytes (TIL) from tumor tissue or from immunocompromised individuals. Further, use of longer incubation times can increase the efficiency of capture of CD8+ T cells. Thus, by simply shortening or lengthening the time T cells are allowed to bind to the CD3/CD28 beads and/or by increasing or decreasing the ratio of beads to T cells (as described further herein), subpopulations of T cells can be preferentially selected for or against at culture initiation or at other time points during the process. Additionally, by increasing or decreasing the ratio of anti-CD3 and/or anti-CD28 antibodies on the beads or other surface, subpopulations of T cells can be preferentially selected for or against at culture initiation or at other desired time points.

In one embodiment, a T cell population can be selected that expresses one or more of IFN-γ, TNFα, IL-17A, IL-2, IL-3, IL-4, GM-CSF, IL-10, IL-13, granzyme B, and perforin, or other appropriate molecules, e.g., other cytokines. Methods for screening for cell expression can be determined, e.g., by the methods described in PCT Publication No.: WO 2013/126712.

For isolation of a desired population of cells by positive or negative selection, the concentration of cells and surface (e.g., particles such as beads) can be varied. In certain aspects, it may be desirable to significantly decrease the volume in which beads and cells are mixed together (e.g., increase the concentration of cells), to ensure maximum contact of cells and beads. For example, in one aspect, a concentration of 10 billion cells/ml, 9 billion/ml, 8 billion/ml, 7 billion/ml, 6 billion/ml, or 5 billion/ml is used. In one aspect, a concentration of 1 billion cells/ml is used. In yet one aspect, a concentration of cells from 75, 80, 85, 90, 95, or 100 million cells/ml is used. In further aspects, concentrations of 125 or 150 million cells/ml can be used.

Using high concentrations can result in increased cell yield, cell activation, and cell expansion. Further, use of high cell concentrations allows more efficient capture of cells that may weakly express target antigens of interest, such as CD28-negative T cells, or from samples where there are many tumor cells present (e.g., leukemic blood, tumor tissue, etc.). Such populations of cells may have therapeutic value and would be desirable to obtain. For example, using high concentration of cells allows more efficient selection of CD8+ T cells that normally have weaker CD28 expression.

In a related aspect, it may be desirable to use lower concentrations of cells. By significantly diluting the mixture of T cells and surface (e.g., particles such as beads), interactions between the particles and cells is minimized. This selects for cells that express high amounts of desired antigens to be bound to the particles. For example, CD4+ T cells express higher levels of CD28 and are more efficiently captured than CD8+ T cells in dilute concentrations. In one aspect, the concentration of cells used is 5×10⁶/ml. In other aspects, the concentration used can be from about 1×10⁵/ml to 1×10⁶/ml, and any integer value in between.

In other aspects, the cells may be incubated on a rotator for varying lengths of time at varying speeds at either 2-10° C. or at room temperature.

T cells for stimulation can also be frozen after a washing step. Wishing not to be bound by theory, the freeze and subsequent thaw step provides a more uniform product by removing granulocytes and to some extent monocytes in the cell population. After the washing step that removes plasma and platelets, the cells may be suspended in a freezing solution. While many freezing solutions and parameters are known in the art and will be useful in this context, one method involves using PBS containing 20% DMSO and 8% human serum albumin, or culture media containing 10% Dextran 40 and 5% Dextrose, 20% Human Serum Albumin and 7.5% DMSO, or 31.25% Plasmalyte-A, 31.25% Dextrose 5%, 0.45% NaCl, 10% Dextran 40 and 5% Dextrose, 20% Human Serum Albumin, and 7.5% DMSO or other suitable cell freezing media containing for example, Hespan and PlasmaLyte A, the cells then are frozen to −80° C. at a rate of 1° per minute and stored in the vapor phase of a liquid nitrogen storage tank. Other methods of controlled freezing may be used as well as uncontrolled freezing immediately at −20° C. or in liquid nitrogen.

In certain aspects, cryopreserved cells are thawed and washed as described herein and allowed to rest for one hour at room temperature prior to activation using the methods of the present disclosure.

Also contemplated in the context of the disclosure is the collection of blood samples or apheresis product from a subject at a time period prior to when the expanded cells as described herein might be needed. As such, the source of the cells to be expanded can be collected at any time point necessary, and desired cells, such as T cells, isolated and frozen for later use in immune effector cell therapy for any number of diseases or conditions that would benefit from immune effector cell therapy, such as those described herein. In one aspect, a blood sample or an apheresis is taken from a generally healthy subject. In certain aspects, a blood sample or an apheresis is taken from a generally healthy subject who is at risk of developing a disease, but who has not yet developed a disease, and the cells of interest are isolated and frozen for later use. In certain aspects, the T cells may be expanded, frozen, and used at a later time. In certain aspects, samples are collected from a patient shortly after diagnosis of a particular disease as described herein but prior to any treatments. In a further aspect, the cells are isolated from a blood sample or an apheresis from a subject prior to any number of relevant treatment modalities, including but not limited to treatment with agents such as natalizumab, efalizumab, antiviral agents, chemotherapy, radiation, immunosuppressive agents, such as cyclosporin, azathioprine, methotrexate, mycophenolate, and FK506, antibodies, or other immunoablative agents such as CAMPATH, anti-CD3 antibodies, cytoxan, fludarabine, cyclosporin, FK506, rapamycin, mycophenolic acid, steroids, FR901228, and irradiation.

In a further aspect of the present disclosure, T cells are obtained from a patient directly following treatment that leaves the subject with functional T cells. In this regard, it has been observed that following certain cancer treatments, in particular treatments with drugs that damage the immune system, shortly after treatment during the period when patients would normally be recovering from the treatment, the quality of T cells obtained may be optimal or improved for their ability to expand ex vivo. Likewise, following ex vivo manipulation using the methods described herein, these cells may be in a preferred state for enhanced engraftment and in vivo expansion. Thus, it is contemplated within the context of the present disclosure to collect blood cells, including T cells, dendritic cells, or other cells of the hematopoietic lineage, during this recovery phase. Further, in certain aspects, mobilization (for example, mobilization with GM-CSF) and conditioning regimens can be used to create a condition in a subject wherein repopulation, recirculation, regeneration, and/or expansion of particular cell types is favored, especially during a defined window of time following therapy. Illustrative cell types include T cells, B cells, dendritic cells, and other cells of the immune system.

In one embodiment, the immune effector cells expressing a CAR molecule, e.g., a CAR molecule described herein, are obtained from a subject that has received a low, immune enhancing dose of an mTOR inhibitor. In an embodiment, the population of immune effector cells, e.g., T cells, to be engineered to express a CAR, are harvested after a sufficient time, or after sufficient dosing of the low, immune enhancing, dose of an mTOR inhibitor, such that the level of PD1 negative immune effector cells, e.g., T cells, or the ratio of PD1 negative immune effector cells, e.g., T cells/PD1 positive immune effector cells, e.g., T cells, in the subject or harvested from the subject has been, at least transiently, increased.

In other embodiments, population of immune effector cells, e.g., T cells, which have, or will be engineered to express a CAR, can be treated ex vivo by contact with an amount of an mTOR inhibitor that increases the number of PD1 negative immune effector cells, e.g., T cells or increases the ratio of PD1 negative immune effector cells, e.g., T cells/PD1 positive immune effector cells, e.g., T cells.

In one embodiment, a T cell population is diaglycerol kinase (DGK)-deficient. DGK-deficient cells include cells that do not express DGK RNA or protein, or have reduced or inhibited DGK activity. DGK-deficient cells can be generated by genetic approaches, e.g., administering RNA-interfering agents, e.g., siRNA, shRNA, miRNA, to reduce or prevent DGK expression. Alternatively, DGK-deficient cells can be generated by treatment with DGK inhibitors described herein.

In one embodiment, a T cell population is Ikaros-deficient. Ikaros-deficient cells include cells that do not express Ikaros RNA or protein, or have reduced or inhibited Ikaros activity, Ikaros-deficient cells can be generated by genetic approaches, e.g., administering RNA-interfering agents, e.g., siRNA, shRNA, miRNA, to reduce or prevent Ikaros expression. Alternatively, Ikaros-deficient cells can be generated by treatment with Ikaros inhibitors, e.g., lenalidomide.

In embodiments, a T cell population is DGK-deficient and Ikaros-deficient, e.g., does not express DGK and Ikaros, or has reduced or inhibited DGK and Ikaros activity. Such DGK and Ikaros-deficient cells can be generated by any of the methods described herein.

In an embodiment, the NK cells are obtained from the subject. In another embodiment, the NK cells are an NK cell line, e.g., NK-92 cell line (Conkwest).

In some aspects, the cells of the disclosure (e.g., the immune effector cells of the disclosure, e.g., the CAR-expressing cells of the disclosure) are induced pluripotent stem cells (“iPSCs”) or embryonic stem cells (ESCs), or are T cells generated from (e.g., differentiated from) said iPSC and/or ESC. iPSCs can be generated, for example, by methods known in the art, from peripheral blood T lymphocytes, e.g., peripheral blood T lymphocytes isolated from a healthy volunteer. As well, such cells may be differentiated into T cells by methods known in the art. See e.g., Themeli M. et al., Nat. Biotechnol., 31, pp. 928-933 (2013); doi: 10.1038/nbt.2678; WO2014/165707, the contents of each of which are incorporated herein by reference in their entirety.

In another embodiment, the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, of the present disclosure are used in combination with one or more of the therapeutic agents listed in Table 13 or listed in the patent and patent applications cited in Table 13, to treat cancer. Each publication listed in Table 13 is herein incorporated by reference in its entirety, including all structural formulae therein.

TABLE 13 Second agent Generic Name Patents/Patent Application No. Tradename Compound Structure Publications A1 Sotrastaurin

EP 16822103 US 2007/142401 WO 2005/039549 A2 Nilotinib HCl monohydrate TASIGNA ®

WO 2004/005281 U.S. Pat. No. 7,169,791 A3

WO2011/023773 A4

WO2012/149413 A6

WO 2010/029082 A7

WO2015/107493 A8 WO2015/107495 A9

WO 2011/076786 A10 Deferasirox EXJADE ®

WO 1997/049395 A11 Letrozole FEMARA ®

U.S. Pat. No. 4,978,672 A12

WO 2013/124826 US 2013/0225574 A13

WO 2013/111105 A14

WO2007/121484 A15 Imatinib mesylate GLEEVEC ®

WO 1999/003854 A16 Capmatinib

EP 2099447 U.S. Pat. No. 7,767,675 U.S. Pat. No. 8,420,645 A17 Ruxolitinib Phosphate JAKAFI ®

WO 2007/070514; EP 2474545 U.S. Pat. No. 7,598,257; WO 2014/018632 A18 Panobinostat

WO 2014/072493 WO 2002/022577 EP 1870399 A20

WO 2008/016893 EP 2051990 U.S. Pat. No. 8,552,003 A21

WO2015/022662 A22 ceritinib ZYKADIA ™

WO 2008/073687 U.S. Pat. No. 8,039,479 A23 Ribociclib KISQALI ®

U.S. Pat. No. 8,415,355 U.S. Pat. No. 8,685,980 A24

WO 2010/007120 A26

WO 2011/101409 A27 Human monoclonal antibody to HER3 WO 2012/022814 EP 2606070 U.S. Pat. No. 8,735,551 A28 Antibody Drug Conjugate (ADC) WO 2014/160160 A29 Monoclonal antibody or Fab to M-CSF WO 2004/045532 A30 Midostaurin

WO 2003/037347 EP 1441737 US 2012/252785 A31 Everolimus AFINITOR ®

WO 1994/009010 WO 2014/085318 A32

WO 2007/030377 U.S. Pat. No. 7,482,367 A34

WO 2006/122806 A35

WO 2008/073687 U.S. Pat. No. 8,372,858 A36 Valspodar AMDRAY ™

EP 296122 A37 Vatalanib succinate

WO 98/35958 A38

WO2014/141104 A39 Asciminib

WO2013/171639 WO2013/171640 WO2013/171641 WO2013/171642 A42

WO2010/015613 WO2013030803 U.S. Pat. No. 7,989,497, A43 WO 2017/025918 WO2011/121418 U.S. Pat. No. 8,796,284 A44

WO2010/101849 A45

WO2014/130310 A46 trametinib

WO2005/121142 U.S. Pat. No. 7,378,423 A47 dabrafenib

WO 2009/137391 U.S. Pat. No. 7,994,185 A49 octreotide

U.S. Pat. No. 4,395,403 EP 0 029 579 A50

WO2016/103155 U.S. Pat. No. 9,580,437 EP 3237418 A51

U.S. Pat. No. 9,512,084 WO/2015/079417 A52

WO 2010/002655 U.S. Pat. No. 8,519,129 A53

WO 2010/002655 U.S. Pat. No. 8,519,129 A54

WO2010/002655 Estrogen Receptor Antagonists

In some embodiments, an estrogen receptor (ER) antagonist is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the estrogen receptor antagonist is a selective estrogen receptor degrader (SERD). SERDs are estrogen receptor antagonists which bind to the receptor and result in e.g., degradation or down-regulation of the receptor (Boer K. et al., (2017) Therapeutic Advances in Medical Oncology 9(7): 465-479). ER is a hormone-activated transcription factor important for e.g., the growth, development and physiology of the human reproductive system. ER is activated by, e.g., the hormone estrogen (17beta estradiol). ER expression and signalling is implicated in cancers (e.g., breast cancer), e.g., ER positive (ER+) breast cancer. In some embodiments, the SERD is chosen from LSZ 102, fulvestrant, brilanestrant, or elacestrant.

Exemplary Estrogen Receptor Antagonists

In some embodiments, the SERD comprises a compound disclosed in International Application Publication No. WO 2014/130310, which is hereby incorporated by reference in its entirety. In some embodiments, the SERD comprises LSZ102. LSZ102 has the chemical name: (E)-3-(4-((2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-hydroxybenzo[b]thiophen-3-yl)oxy)phenyl)acrylic acid.

Other Exemplary Estrogen Receptor Antagonists

In some embodiments, the SERD comprises fulvestrant (CAS Registry Number: 129453-61-8), or a compound disclosed in International Application Publication No. WO 2001/051056, which is hereby incorporated by reference in its entirety. Fulvestrant is also known as ICI 182780, ZM 182780, FASLODEX®, or (7a,17(3)-7-{9-[(4,4,5,5,5-pentafluoropentyl)sulfinyl]nonyl}estra-1,3,5(10)-triene-3,17-diol. Fulvestrant is a high affinity estrogen receptor antagonist with an IC50 of 0.29 nM.

In some embodiments, the SERD comprises elacestrant (CAS Registry Number: 722533-56-4), or a compound disclosed in U.S. Pat. No. 7,612,114, which is incorporated by reference in its entirety. Elacestrant is also known as RAD1901, ER-306323 or (6R)-6-{2-[Ethyl({4-[2-(ethylamino)ethyl]phenyl}methyl)amino]-4-methoxyphenyl}-5,6,7,8-tetrahydronaphthalen-2-ol. Elacestrant is an orally bioavailable, non-steroidal combined selective estrogens receptor modulator (SERM) and a SERD. Elacestrant is also disclosed, e.g., in Garner F et al., (2015) Anticancer Drugs 26(9):948-56.

In some embodiments, the SERD is brilanestrant (CAS Registry Number: 1365888-06-7), or a compound disclosed in International Application Publication No. WO 2015/136017, which is incorporated by reference in its entirety. Brilanestrant is also known as GDC-0810, ARN810, RG-6046, RO-7056118 or (2E)-3-{4-[(1E)-2-(2-chloro-4-fluorophenyl)-1-(1H-indazol-5-yl)but-1-en-1-yl]phenyl}prop-2-enoic acid. Brilanestrant is a next-generation, orally bioavailable selective SERD with an IC50 of 0.7 nM. Brilanestrant is also disclosed, e.g., in Lai A. et al. (2015) Journal of Medicinal Chemistry 58 (12): 4888-4904.

In some embodiments, the SERD is chosen from RU 58668, GW7604, AZD9496, bazedoxifene, pipendoxifene, arzoxifene, OP-1074, or acolbifene, e.g., as disclosed in McDonell et al. (2015) Journal of Medicinal Chemistry 58(12) 4883-4887. Other exemplary estrogen receptor antagonists are disclosed, e.g., in WO 2011/156518, WO 2011/159769, WO 2012/037410, WO 2012/037411, and US 2012/0071535, all of which are hereby incorporated by reference in their entirety.

CDK4/6 Inhibitors

In some embodiments, an inhibitor of Cyclin-Dependent Kinases 4 or 6 (CDK4/6) is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the CDK4/6 inhibitor is chosen from ribociclib, abemaciclib (Eli Lilly), or palbociclib.

Exemplary CDK4/6 Inhibitors

In some embodiments, the CDK4/6 inhibitor comprises ribociclib (CAS Registry Number: 1211441-98-3), or a compound disclosed in U.S. Pat. Nos. 8,415,355 and 8,685,980, which are incorporated by reference in their entirety.

In some embodiments, the CDK4/6 inhibitor comprises a compound disclosed in International Application Publication No. WO 2010/020675 and U.S. Pat. Nos. 8,415,355 and 8,685,980, which are incorporated by reference in their entirety.

In some embodiments, the CDK4/6 inhibitor comprises ribociclib (CAS Registry Number: 1211441-98-3). Ribociclib is also known as LEE011, KISQALI®, or 7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide.

Other Exemplary CDK46 Inhibitors

In some embodiments, the CDK4/6 inhibitor comprises abemaciclib (CAS Registry Number: 1231929-97-7). Abemaciclib is also known as LY835219 or N-[5-[(4-Ethyl-1-piperazinyl)methyl]-2-pyridinyl]-5-fluoro-4-[4-fluoro-2-methyl-1-(1-methylethyl)-1H-benzimidazol-6-yl]-2-pyrimidinamine. Abemaciclib is a CDK inhibitor selective for CDK4 and CDK6 and is disclosed, e.g., in Torres-Guzman R et al. (2017) Oncotarget 10.18632/oncotarget. 17778.

In some embodiments, the CDK4/6 inhibitor comprises palbociclib (CAS Registry Number: 571190-30-2). Palbociclib is also known as PD-0332991, IBRANCE® or 6-Acetyl-8-cyclopentyl-5-methyl-2-{[5-(1-piperazinyl)-2-pyridinyl]amino}pyrido[2,3-d]pyrimidin-7(8H)-one. Palbociclib inhibits CDK4 with an IC50 of 11 nM, and inhibits CDK6 with an IC50 of 16 nM, and is disclosed, e.g., in Finn et al. (2009) Breast Cancer Research 11(5):R77.

CXCR2 Inhibitors

In some embodiments, an inhibitor of chemokine (C-X-C motif) receptor 2 (CXCR2) is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the CXCR2 inhibitor is chosen from 6-chloro-3-((3,4-dioxo-2-(pentan-3-ylamino)cyclobut-1-en-1-yl)amino)-2-hydroxy-N-methoxy-N-methylbenzenesulfonamide, danirixin, reparixin, or navarixin.

Exemplary CXCR2 Inhibitors

In some embodiments, the CXCR2 inhibitor comprises a compound disclosed in U.S. Pat. Nos. 7,989,497, 8,288,588, 8,329,754, 8,722,925, 9,115,087, U.S. Application Publication Nos. US 2010/0152205, US 2011/0251205 and US 2011/0251206, and International Application Publication Nos. WO 2008/061740, WO 2008/061741, WO 2008/062026, WO 2009/106539, WO2010/063802, WO 2012/062713, WO 2013/168108, WO 2010/015613 and WO 2013/030803. In some embodiments, the CXCR2 inhibitor comprises 6-chloro-3-((3,4-dioxo-2-(pentan-3-ylamino)cyclobut-1-en-1-yl)amino)-2-hydroxy-N-methoxy-N-methylbenzenesulfonamide or a choline salt thereof. In some embodiments, the CXCR2 inhibitor comprises 6-chloro-3-((3,4-dioxo-2-(pentan-3-ylamino)cyclobut-1-en-1-yl)amino)-2-hydroxy-N-methoxy-N-methylbenzenesulfonamide choline salt. In some embodiments, the CXCR2 inhibitor is 2-Hydroxy-N,N,N-trimethylethan-1-aminium 3-chloro-6-({3,4-dioxo-2-[(pentan-3-yl)amino]cyclobut-1-en-1-yl}amino)-2-(N-methoxy-N-methylsulfamoyl)phenolate (i.e., 6-chloro-3-((3,4-dioxo-2-(pentan-3-ylamino)cyclobut-1-en-1-yl)amino)-2-hydroxy-N-methoxy-N-methylbenzenesulfonamide choline salt) and has the following chemical structure:

Other Exemplary CXCR2 Inhibitors

In some embodiments, the CXCR2 inhibitor comprises danirixin (CAS Registry Number: 954126-98-8). Danirixin is also known as GSK1325756 or 1-(4-chloro-2-hydroxy-3-piperidin-3-ylsulfonylphenyl)-3-(3-fluoro-2-methylphenyl)urea. Danirixin is disclosed, e.g., in Miller et al. Eur J Drug Metab Pharmacokinet (2014) 39:173-181; and Miller et al. BMC Pharmacology and Toxicology (2015), 16:18.

In some embodiments, the CXCR2 inhibitor comprises reparixin (CAS Registry Number: 266359-83-5). Reparixin is also known as repertaxin or (2R)-2-[4-(2-methylpropyl)phenyl]-N-methylsulfonylpropanamide. Reparixin is a non-competitive allosteric inhibitor of CXCR1/2. Reparixin is disclosed, e.g., in Zarbock et al. Br J Pharmacol. 2008; 155(3):357-64.

In some embodiments, the CXCR2 inhibitor comprises navarixin. Navarixin is also known as MK-7123, SCH 527123, PS291822, or 2-hydroxy-N,N-dimethyl-3-[[2-[[(1R)-1-(5-methylfuran-2-yl)propyl]amino]-3,4-dioxocyclobuten-1-yl]amino]benzamide. Navarixin is disclosed, e.g., in Ning et al. Mol Cancer Ther. 2012; 11(6):1353-64.

CSF-1/1R Binding Agents

In some embodiments, a CSF-1/1R binding agent is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the CSF-1/1R binding agent is chosen from an inhibitor of macrophage colony-stimulating factor (M-CSF), e.g., a monoclonal antibody or Fab to M-CSF (e.g., MCS110), a CSF-1R tyrosine kinase inhibitor (e.g., 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide or BLZ945), a receptor tyrosine kinase inhibitor (RTK) (e.g., pexidartinib), or an antibody targeting CSF-1R (e.g., emactuzumab or FPA008). In some embodiments, the CSF-1/1R inhibitor is BLZ945. In some embodiments, the CSF-1/1R binding agent is MCS110. In other embodiments, the CSF-1/1R binding agent is pexidartinib.

Exemplary CSF-1 Binding Agents

In some embodiments, the CSF-1/1R binding agent comprises an inhibitor of macrophage colony-stimulating factor (M-CSF). M-CSF is also sometimes known as CSF-1. In certain embodiments, the CSF-1/1R binding agent is an antibody to CSF-1 (e.g., MCS110). In other embodiments, the CSF-1/1R binding agent is an inhibitor of CSF-1R (e.g., BLZ945).

In some embodiments, the CSF-1/1R binding agent comprises a monoclonal antibody or Fab to M-CSF (e.g., MCS110/H-RX1), or a binding agent to CSF-1 disclosed in International Application Publication Nos. WO 2004/045532 and WO 2005/068503, including H-RX1 or 5H4 (e.g., an antibody molecule or Fab fragment against M-CSF) and U.S. Pat. No. 9,079,956, which applications and patent are incorporated by reference in their entirety.

TABLE 13a Amino acid and nucleotide sequences of an exemplary anti-M-CSF antibody molecule (MCS110) (H-RX1) HC QVQLQESGPGLVKPSQTLSLTCTVSDYSITSDYAWNWIRQFPGKGLEWMG YISYSGSTSYNPSLKSRITISRDTSKNQFSLQLNSVTAADTAVYYCASFDYA HAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 271) (H-RX1) LC DIVLTQSPAFLSVTPGEKVTFTCQASQSIGTSIHWYQQKTDQAPKLLIKYAS ESISGIPSRFSGSGSGTDFTLTISSVEAEDAADYYCQQINSWPTTFGGGTKLEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQS GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC (SEQ ID NO: 272) Heavy Chain SDYAWN (SEQ ID NO: 273) CDR1 (Kabat) Heavy Chain YISYSGSTSYNPSLKS (SEQ ID NO: 274) CDR2 (Kabat) Heavy Chain FDYAHAMDY (SEQ ID NO: 275) CDR3 (Kabat) Light Chain QASQSIGTSIH (SEQ ID NO: 276) CDR1 (Kabat) Light Chain YASESIS (SEQ ID NO: 277) CDR2 (Kabat) Light Chain QQINSWPTT (SEQ ID NO: 278) CDR3 (Kabat)

In another embodiment, the CSF-1/1R binding agent comprises a CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (BLZ945), or a compound disclosed in International Application Publication No. WO 2007/121484, and U.S. Pat. Nos. 7,553,854, 8,173,689, and 8,710,048, which are incorporated by reference in their entirety.

Other Exemplary CSF-1/1R Binding Agents

In some embodiments, the CSF-1/1R binding agent comprises pexidartinib (CAS Registry Number 1029044-16-3). Pexidrtinib is also known as PLX3397 or 5-((5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)pyridin-2-amine. Pexidartinib is a small-molecule receptor tyrosine kinase (RTK) inhibitor of KIT, CSF1R and FLT3. FLT3, CSF1R and FLT3 are overexpressed or mutated in many cancer cell types and play major roles in tumor cell proliferation and metastasis. PLX3397 can bind to and inhibit phosphorylation of stem cell factor receptor (KIT), colony-stimulating factor-1 receptor (CSF1R) and FMS-like tyrosine kinase 3 (FLT3), which may result in the inhibition of tumor cell proliferation and down-modulation of macrophages, osteoclasts and mast cells involved in the osteolytic metastatic disease.

In some embodiments, the CSF-1/1R binding agent is emactuzumab. Emactuzumab is also known as RG7155 or RO5509554. Emactuzumab is a humanized IgG1 mAb targeting CSF1R. In some embodiments, the CSF-1/1R binding agent is FPA008. FPA008 is a humanized mAb that inhibits CSF1R.

A2aR Antagonists

In some embodiments, an adenosine A2a receptor (A2aR) antagonist (e.g., an inhibitor of A2aR pathway, e.g., an adenosine inhibitor, e.g., an inhibitor of A2aR or CD-73) is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the A2aR antagonist is selected from PBF509 (NIR178) (Palobiofarma/Novartis), CPI444/V81444 (Corvus/Genentech), AZD4635/HTL-1071 (AstraZeneca/Heptares), Vipadenant (Redox/Juno), GBV-2034 (Globavir), AB928 (Arcus Biosciences), Theophylline, Istradefylline (Kyowa Hakko Kogyo), Tozadenant/SYN-115 (Acorda), KW-6356 (Kyowa Hakko Kogyo), ST-4206 (Leadiant Biosciences), and Preladenant/SCH 420814 (Merck/Schering).

Exemplary A2aR Antagonists

In some embodiments, the A2aR antagonist comprises PBF509 (NIR178) or a compound disclosed in U.S. Pat. No. 8,796,284 or in International Application Publication No. WO 2017/025918, herein incorporated by reference in their entirety. PBF509 (NIR178) is also known as NIR178.

Other Exemplary A2aR Antagonists

In certain embodiments, the A2aR antagonist comprises CPI444/V81444. CPI-444 and other A2aR antagonists are disclosed in International Application Publication No. WO 2009/156737, herein incorporated by reference in its entirety. In certain embodiments, the A2aR antagonist is (S)-7-(5-methylfuran-2-yl)-3-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-5-amine. In certain embodiments, the A2aR antagonist is (R)-7-(5-methylfuran-2-yl)-3-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-5-amine, or racemate thereof. In certain embodiments, the A2aR antagonist is 7-(5-methylfuran-2-yl)-3-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-5-amine.

In certain embodiments, the A2aR antagonist is AZD4635/HTL-1071. A2aR antagonists are disclosed in International Application Publication No. WO 2011/095625, herein incorporated by reference in its entirety. In certain embodiments, the A2aR antagonist is 6-(2-chloro-6-methylpyridin-4-yl)-5-(4-fluorophenyl)-1,2,4-triazin-3-amine.

In certain embodiments, the A2aR antagonist is ST-4206 (Leadiant Biosciences). In certain embodiments, the A2aR antagonist is an A2aR antagonist described in U.S. Pat. No. 9,133,197, herein incorporated by reference in its entirety.

In certain embodiments, the A2aR antagonist is an A2aR antagonist described in U.S. Pat. Nos. 8,114,845 and 9,029,393, U.S. Application Publication Nos. 2017/0015758 and 2016/0129108, herein incorporated by reference in their entirety.

In some embodiments, the A2aR antagonist is istradefylline (CAS Registry Number: 155270-99-8). Istradefylline is also known as KW-6002 or 8-[(E)-2-(3,4-dimethoxyphenyl)vinyl]-1,3-diethyl-7-methyl-3,7-dihydro-1H-purine-2,6-dione. Istradefylline is disclosed, e.g., in LeWitt et al. (2008) Annals of Neurology 63 (3): 295-302).

In some embodiments, the A2aR antagonist is tozadenant (Biotie). Tozadenant is also known as SYN115 or 4-hydroxy-N-(4-methoxy-7-morpholin-4-yl-1,3-benzothiazol-2-yl)-4-methylpiperidine-1-carboxamide. Tozadenant blocks the effect of endogenous adenosine at the A2a receptors, resulting in the potentiation of the effect of dopamine at the D2 receptor and inhibition of the effect of glutamate at the mGluR5 receptor. In some embodiments, the A2aR antagonist is preladenant (CAS Registry Number: 377727-87-2). Preladenant is also known as SCH 420814 or 2-(2-Furanyl)-7-[2-[4-[4-(2-methoxyethoxy)phenyl]-1-piperazinyl]ethyl]7H-pyrazolo[4,3-e][1,2,4]triazolo[1,5-c]pyrimidine-5-amine. Preladenant was developed as a drug that acted as a potent and selective antagonist at the adenosine A2A receptor.

In some embodiments, the A2aR antagonist is vipadenan. Vipadenan is also known as BIIB014, V2006, or 3-[(4-amino-3-methylphenyl)methyl]-7-(furan-2-yl)triazolo[4,5-d]pyrimidin-5-amine. Other exemplary A2aR antagonists include, e.g., ATL-444, MSX-3, SCH-58261, SCH-412,348, SCH-442,416, VER-6623, VER-6947, VER-7835, CGS-15943, and ZM-241,385.

In some embodiments, the A2aR antagonist is an A2aR pathway antagonist (e.g., a CD-73 inhibitor, e.g., an anti-CD73 antibody) is MEDI9447. MEDI9447 is a monoclonal antibody specific for CD73. Targeting the extracellular production of adenosine by CD73 may reduce the immunosuppressive effects of adenosine. MEDI9447 was reported to have a range of activities, e.g., inhibition of CD73 ectonucleotidase activity, relief from AMP-mediated lymphocyte suppression, and inhibition of syngeneic tumor growth. MEDI9447 can drive changes in both myeloid and lymphoid infiltrating leukocyte populations within the tumor microenvironment. These changes include, e.g., increases in CD8 effector cells and activated macrophages, as well as a reduction in the proportions of myeloid-derived suppressor cells (MDSC) and regulatory T lymphocytes.

IDO Inhibitors

In some embodiments, an inhibitor of indoleamine 2,3-dioxygenase (IDO) and/or tryptophan 2,3-dioxygenase (TDO) is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the IDO inhibitor is chosen from (4E)-4-[(3-chloro-4-fluoroanilino)-nitrosomethylidene]-1,2,5-oxadiazol-3-amine (also known as epacadostat or INCB24360), indoximod ( ), (1-methyl-D-tryptophan), α-cyclohexyl-5H-Imidazo[5,1-a]isoindole-5-ethanol (also known as NLG919), indoximod, and BMS-986205 (formerly F001287).

Exemplary IDO Inhibitors

In some embodiments, the IDO/TDO inhibitor is indoximod (New Link Genetics). Indoximod, the D isomer of 1-methyl-tryptophan, is an orally administered small-molecule indoleamine 2,3-dioxygenase (IDO) pathway inhibitor that disrupts the mechanisms by which tumors evade immune-mediated destruction.

In some embodiments, the IDO/TDO inhibitor is NLG919 (New Link Genetics). NLG919 is a potent IDO (indoleamine-(2,3)-dioxygenase) pathway inhibitor with Ki/EC50 of 7 nM/75 nM in cell-free assays.

In some embodiments, the IDO/TDO inhibitor is epacadostat (CAS Registry Number: 1204669-58-8). Epacadostat is also known as INCB24360 or INCB024360 (Incyte). Epacadostat is a potent and selective indoleamine 2,3-dioxygenase (IDO1) inhibitor with IC50 of 10 nM, highly selective over other related enzymes such as IDO2 or tryptophan 2,3-dioxygenase (TDO).

In some embodiments, the IDO/TDO inhibitor is F001287 (Flexus/BMS). F001287 is a small molecule inhibitor of indoleamine 2,3-dioxygenase 1 (IDO1).

STING Agonists

In some embodiments, a STING agonist is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the STING agonist is cyclic dinucleotide, e.g., a cyclic dinucleotide comprising purine or pyrimidine nucleobases (e.g., adenosine, guanine, uracil, thymine, or cytosine nucleobases). In some embodiments, the nucleobases of the cyclic dinucleotide comprise the same nucleobase or different nucleobases.

In some embodiments, the STING agonist comprises an adenosine or a guanosine nucleobase. In some embodiments, the STING agonist comprises one adenosine nucleobase and one guanosine nucleobase. In some embodiments, the STING agonist comprises two adenosine nucleobases or two guanosine nucleobases.

In some embodiments, the STING agonist comprises a modified cyclic dinucleotide, e.g., comprising a modified nucleobase, a modified ribose, or a modified phosphate linkage. In some embodiments, the modified cyclic dinucleotide comprises a modified phosphate linkage, e.g., a thiophosphate.

In some embodiments, the STING agonist comprises a cyclic dinucleotide (e.g., a modified cyclic dinucleotide) with 2′,5′ or 3′,5′ phosphate linkages. In some embodiments, the STING agonist comprises a cyclic dinucleotide (e.g., a modified cyclic dinucleotide) with Rp or Sp stereochemistry around the phosphate linkages.

In some embodiments, the STING agonist is MK-1454 (Merck). MK-1454 is a cyclic dinucleotide Stimulator of Interferon Genes (STING) agonist that activates the STING pathway. Exemplary STING agonist are disclosed, e.g., in PCT Publication No. WO 2017/027645.

Galectin Inhibitors

In some embodiments, a Galectin, e.g., Galectin-1 or Galectin-3, inhibitor is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the combination comprises a Galectin-1 inhibitor and a Galectin-3 inhibitor. In some embodiments, the combination comprises a bispecific inhibitor (e.g., a bispecific antibody molecule) targeting both Galectin-1 and Galectin-3. In some embodiments, the Galectin inhibitor is chosen from an anti-Galectin antibody molecule, GR-MD-02 (Galectin Therapeutics), Galectin-3C (Mandal Med), Anginex, or OTX-008 (OncoEthix, Merck). Galectins are a family of proteins that bind to beta galactosidase sugars.

The Galectin family of proteins comprises at least of Galectin-1, Galectin-2, Galectin-3, Galectin-4, Galectin-7, and Galectin-8. Galectins are also referred to as S-type lectins, and are soluble proteins with, e.g., intracellular and extracellular functions.

Galectin-1 and Galectin-3 are highly expressed in various tumor types. Galectin-1 and Galectin-3 can promote angiogenesis and/or reprogram myeloid cells toward a pro-tumor phenotype, e.g., enhance immunosuppression from myeloid cells. Soluble Galectin-3 can also bind to and/or inactivate infiltrating T cells.

Exemplary Galectin Inhibitors

In some embodiments, a Galectin inhibitor is an antibody molecule. In an embodiment, an antibody molecule is a monospecific antibody molecule and binds a single epitope. E.g., a monospecific antibody molecule having a plurality of immunoglobulin variable domain sequences, each of which binds the same epitope. In an embodiment, the Galectin inhibitor is an anti-Galectin, e.g., anti-Galectin-1 or anti-Galectin-3, antibody molecule. In some embodiments, the Galectin inhibitor is an anti-Galectin-1 antibody molecule. In some embodiments, the Galectin inhibitor is an anti-Galectin-3 antibody molecule.

In an embodiment an antibody molecule is a multispecific antibody molecule, e.g., it comprises a plurality of immunoglobulin variable domains sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In an embodiment, the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein). In an embodiment, the first and second epitopes overlap. In an embodiment, the first and second epitopes do not overlap. In an embodiment, the first and second epitopes are on different antigens, e.g., the different proteins (or different subunits of a multimeric protein). In an embodiment, a multispecific antibody molecule comprises a third, fourth or fifth immunoglobulin variable domain. In an embodiment, a multispecific antibody molecule is a bispecific antibody molecule, a trispecific antibody molecule, or tetraspecific antibody molecule.

In an embodiment, the Galectin inhibitor is a multispecific antibody molecule. In an embodiment, a multispecific antibody molecule is a bispecific antibody molecule. A bispecific antibody has specificity for no more than two antigens. A bispecific antibody molecule is characterized by a first immunoglobulin variable domain sequence which has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope. In an embodiment, the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein). In an embodiment, the first and second epitopes overlap. In an embodiment, the first and second epitopes do not overlap. In an embodiment, the first and second epitopes are on different antigens, e.g., the different proteins (or different subunits of a multimeric protein). In an embodiment a bispecific antibody molecule comprises a heavy chain variable domain sequence and a light chain variable domain sequence which have binding specificity for a first epitope and a heavy chain variable domain sequence and a light chain variable domain sequence which have binding specificity for a second epitope. In an embodiment, a bispecific antibody molecule comprises a half antibody having binding specificity for a first epitope and a half antibody having binding specificity for a second epitope. In an embodiment, a bispecific antibody molecule comprises a half antibody, or fragment thereof, having binding specificity for a first epitope and a half antibody, or fragment thereof, having binding specificity for a second epitope. In an embodiment, a bispecific antibody molecule comprises a scFv, or fragment thereof, have binding specificity for a first epitope and a scFv, or fragment thereof, have binding specificity for a second epitope. In an embodiment, the Galectin inhibitor is a bispecific antibody molecule. In an embodiment, the first epitope is located on Galectin-1, and the second epitope is located on Galectin-3.

Protocols for generating bispecific or heterodimeric antibody molecules are known in the art; including but not limited to, for example, the “knob in a hole” approach described in, e.g., U.S. Pat. No. 5,731,168; the electrostatic steering Fc pairing as described in, e.g., WO 09/089004, WO 06/106905 and WO 2010/129304; Strand Exchange Engineered Domains (SEED) heterodimer formation as described in, e.g., WO 07/110205; Fab arm exchange as described in, e.g., WO 08/119353, WO 2011/131746, and WO 2013/060867; double antibody conjugate, e.g., by antibody cross-linking to generate a bi-specific structure using a heterobifunctional reagent having an amine-reactive group and a sulfhydryl reactive group as described in, e.g., U.S. Pat. No. 4,433,059; bispecific antibody determinants generated by recombining half antibodies (heavy-light chain pairs or Fabs) from different antibodies through cycle of reduction and oxidation of disulfide bonds between the two heavy chains, as described in, e.g., U.S. Pat. No. 4,444,878; trifunctional antibodies, e.g., three Fab′ fragments cross-linked through sulfhydryl reactive groups, as described in, e.g., U.S. Pat. No. 5,273,743; biosynthetic binding proteins, e.g., pair of scFvs cross-linked through C-terminal tails preferably through disulfide or amine-reactive chemical cross-linking, as described in, e.g., U.S. Pat. No. 5,534,254; bifunctional antibodies, e.g., Fab fragments with different binding specificities dimerized through leucine zippers (e.g., c-fos and c-jun) that have replaced the constant domain, as described in, e.g., U.S. Pat. No. 5,582,996; bispecific and oligospecific mono- and oligovalent receptors, e.g., VH-CH1 regions of two antibodies (two Fab fragments) linked through a polypeptide spacer between the CH1 region of one antibody and the VH region of the other antibody typically with associated light chains, as described in, e.g., U.S. Pat. No. 5,591,828; bispecific DNA-antibody conjugates, e.g., crosslinking of antibodies or Fab fragments through a double stranded piece of DNA, as described in, e.g., U.S. Pat. No. 5,635,602; bispecific fusion proteins, e.g., an expression construct containing two scFvs with a hydrophilic helical peptide linker between them and a full constant region, as described in, e.g., U.S. Pat. No. 5,637,481; multivalent and multispecific binding proteins, e.g., dimer of polypeptides having first domain with binding region of Ig heavy chain variable region, and second domain with binding region of Ig light chain variable region, generally termed diabodies (higher order structures are also disclosed creating bispecific, trispecific, or tetraspecific molecules, as described in, e.g., U.S. Pat. No. 5,837,242; minibody constructs with linked VL and VH chains further connected with peptide spacers to an antibody hinge region and CH3 region, which can be dimerized to form bispecific/multivalent molecules, as described in, e.g., U.S. Pat. No. 5,837,821; VH and VL domains linked with a short peptide linker (e.g., 5 or 10 amino acids) or no linker at all in either orientation, which can form dimers to form bispecific diabodies; trimers and tetramers, as described in, e.g., U.S. Pat. No. 5,844,094; String of VH domains (or VL domains in family members) connected by peptide linkages with crosslinkable groups at the C-terminus further associated with VL domains to form a series of FVs (or scFvs), as described in, e.g., U.S. Pat. No. 5,864,019; and single chain binding polypeptides with both a VH and a VL domain linked through a peptide linker are combined into multivalent structures through non-covalent or chemical crosslinking to form, e.g., homobivalent, heterobivalent, trivalent, and tetravalent structures using both scFV or diabody type format, as described in, e.g., U.S. Pat. No. 5,869,620. Additional exemplary multispecific and bispecific molecules and methods of making the same are found, for example, in U.S. Pat. Nos. 5,910,573, 5,932,448, 5,959,083, 5,989,830, 6,005,079, 6,239,259, 6,294,353, 6,333,396, 6,476,198, 6,511,663, 6,670,453, 6,743,896, 6,809,185, 6,833,441, 7,129,330, 7,183,076, 7,521,056, 7,527,787, 7,534,866, 7,612,181, US2002/004587A1, US2002/076406A1, US2002/103345A1, US2003/207346A1, US2003/211078A1, US2004/219643A1, US2004/220388A1, US2004/242847A1, US2005/003403A1, US2005/004352A1, US2005/069552A1, US2005/079170A1, US2005/100543A1, US2005/136049A1, US2005/136051A1, US2005/163782A1, US2005/266425A1, US2006/083747A1, US2006/120960A1, US2006/204493A1, US2006/263367A1, US2007/004909A1, US2007/087381A1, US2007/128150A1, US2007/141049A1, US2007/154901A1, US2007/274985A1, US2008/050370A1, US2008/069820A1, US2008/152645A1, US2008/171855A1, US2008/241884A1, US2008/254512A1, US2008/260738A1, US2009/130106A1, US2009/148905A1, US2009/155275A1, US2009/162359A1, US2009/162360A1, US2009/175851A1, US2009/175867A1, US2009/232811A1, US2009/234105A1, US2009/263392A1, US2009/274649A1, EP346087A2, WO00/06605A2, WO02/072635A2, WO04/081051A1, WO06/020258A2, WO2007/044887A2, WO2007/095338A2, WO2007/137760A2, WO2008/119353A1, WO2009/021754A2, WO2009/068630A1, WO91/03493A1, WO93/23537A1, WO94/09131A1, WO94/12625A2, WO95/09917A1, WO96/37621A2, WO99/64460A1. The contents of the above-referenced applications are incorporated herein by reference in their entireties.

In other embodiments, the anti-Galectin, e.g., anti-Galectin-1 or anti-Galectin-3, antibody molecule (e.g., a monospecific, bispecific, or multispecific antibody molecule) is covalently linked, e.g., fused, to another partner e.g., a protein, e.g., as a fusion molecule for example a fusion protein. In one embodiment, a bispecific antibody molecule has a first binding specificity to a first target (e.g., to Galectin-1), a second binding specificity to a second target (e.g., Galectin-3).

This invention provides an isolated nucleic acid molecule encoding the above antibody molecule, vectors and host cells thereof. The nucleic acid molecule includes but is not limited to RNA, genomic DNA and cDNA.

In some embodiments, a Galectin inhibitor is a peptide, e.g., protein, which can bind to, and inhibit Galectin, e.g., Galectin-1 or Galectin-3, function. In some embodiments, the Galectin inhibitor is a peptide which can bind to, and inhibit Galectin-3 function. In some embodiments, the Galectin inhibitor is the peptide Galectin-3C. In some embodiments, the Galectin inhibitor is a Galectin-3 inhibitor disclosed in U.S. Pat. No. 6,770,622, which is hereby incorporated by reference in its entirety.

Galectin-3C is an N-terminal truncated protein of Galectin-3, and functions, e.g., as a competitive inhibitor of Galectin-3. Galectin-3C prevents binding of endogenous Galectin-3 to e.g., laminin on the surface of, e.g., cancer cells, and other beta-galactosidase glycoconjugates in the extracellular matrix (ECM). Galectin-3C and other exemplary Galectin inhibiting peptides are disclosed in U.S. Pat. No. 6,770,622.

In some embodiments, Galectin-3C comprises the amino acid sequence of SEQ ID NO: 279, or an amino acid substantially identical (e.g., 90, 95 or 99%) identical thereto.

(SEQ ID NO: 279) GAPAGPLIVPYNLPLPGGVVPRMLITILGTVKPNANRIALDFQRGND VAFHFNPRFNENNRRVIVCNTKLDNNWGREERQSVFPFESGKPFKIQVL VEPDHFKVAVNDAHLLQYNHRVKKLNEISKLGISGDIDITSASYTMI.

In some embodiments, the Galectin inhibitor is a peptide, which can bind to, and inhibit Galectin-1 function. In some embodiments, the Galectin inhibitor is the peptide Anginex: Anginex is an anti-angiongenic peptide that binds Galectin-1 (Salomonsson E, et al., (2011) Journal of Biological Chemistry, 286(16):13801-13804). Binding of Anginex to Galectin-1 can interfere with, e.g., the pro-angiongenic effects of Galectin-1.

In some embodiments, the Galectin, e.g., Galectin-1 or Galectin-3, inhibitor is a non-peptidic topomimetic molecule. In some embodiments, the non-peptidic topomimetic Galectin inhibitor is OTX-008 (OncoEthix). In some embodiments, the non-peptidic topomimetic is a non-peptidic topomimetic disclosed in U.S. Pat. No. 8,207,228, which is herein incorporated by reference in its entirety. OTX-008, also known as PTX-008 or Calixarene 0118, is a selective allosteric inhibitor of Galectin-1. OTX-008 has the chemical name: N-[2-(dimethylamino)ethyl]-2-{[26,27,28-tris({[2-(dimethylamino)ethyl]carbamoyl}methoxy) pentacyclo[19.3.1.1,7.1,15,]octacosa-1(25),3(28),4,6,9(27), 1012,15,17,19(26),21,23-dodecaen-25-yl]oxy}acetamide.

In some embodiments, the Galectin, e.g., Galectin-1 or Galectin-3, inhibitor is a carbohydrate based compound. In some embodiments, the Galectin inhibitor is GR-MD-02 (Galectin Therapeutics).

In some embodiments, GR-MD-02 is a Galectin-3 inhibitor. GR-MD-02 is a galactose-pronged polysaccharide also referred to as, e.g., a galactoarabino-rhamnogalaturonate. GR-MD-02 and other galactose-pronged polymers, e.g., galactoarabino-rhamnogalaturonates, are disclosed in U.S. Pat. No. 8,236,780 and U.S. Publication 2014/0086932, the entire contents of which are herein incorporated by reference in their entirety. MEK Inhibitors

In some embodiments, a MEK inhibitor is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the MEK inhibitor is chosen from Trametinib, selumetinib, AS703026, BIX 02189, BIX 02188, CI-1040, PD0325901, PD98059, U0126, XL-518, G-38963, or G02443714. In some embodiments, the MEK inhibitor is Trametinib.

Exemplary MEK Inhibitors

In some embodiments, the MEK inhibitor is trametinib. Trametinib is also known as JTP-74057, TMT212, N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H)-yl}phenyl)acetamide, or Mekinist (CAS Number 871700-17-3).

Other Exemplary MEK Inhibitors

In some embodiments the MEK inhibitor comprises selumetinib which has the chemical name: (5-[(4-bromo-2-chlorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6-carboxamide. Selumetinib is also known as AZD6244 or ARRY 142886, e.g., as described in PCT Publication No. WO2003077914.

In some embodiments, the MEK inhibitor comprises AS703026, BIX 02189 or BIX 02188.

In some embodiments, the MEK inhibitor comprises 2-[(2-Chloro-4-iodophenyl)amino]-N-(cyclopropylmethoxy)-3,4-difluoro-benzamide (also known as CI-1040 or PD184352), e.g., as described in PCT Publication No. WO2000035436).

In some embodiments, the MEK inhibitor comprises N-[(2R)-2,3-Dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide (also known as PD0325901), e.g., as described in PCT Publication No. WO2002006213).

In some embodiments, the MEK inhibitor comprises 2′-amino-3′-methoxyflavone (also known as PD98059) which is available from Biaffin GmbH & Co., KG, Germany.

In some embodiments, the MEK inhibitor comprises 2,3-bis[amino[(2-aminophenyl)thio]methylene]-butanedinitrile (also known as U0126), e.g., as described in U.S. Pat. No. 2,779,780).

In some embodiments, the MEK inhibitor comprises XL-518 (also known as GDC-0973) which has a CAS No. 1029872-29-4 and is available from ACC Corp.

In some embodiments, the MEK inhibitor comprises G-38963.

In some embodiments, the MEK inhibitor comprises G02443714 (also known as AS703206)

Additional examples of MEK inhibitors are disclosed in WO 2013/019906, WO 03/077914, WO 2005/121142, WO 2007/04415, WO 2008/024725 and WO 2009/085983, the contents of which are incorporated herein by reference. Further examples of MEK inhibitors include, but are not limited to, 2,3-Bis[amino[(2-aminophenyl)thio]methylene]-butanedinitrile (also known as U0126 and described in U.S. Pat. No. 2,779,780); (3S,4R,5Z,8S,9S,11E)-14-(Ethylamino)-8,9,16-trihydroxy-3,4-dimethyl-3,4,9, 19-tetrahydro-1H-2-benzoxacyclotetradecine-1,7(8H)-dione] (also known as E6201, described in PCT Publication No. WO2003076424); vemurafenib (PLX-4032, CAS 918504-65-1); (R)-3-(2,3-Dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733, CAS 1035555-63-5); pimasertib (AS-703026, CAS 1204531-26-9); 2-(2-Fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide (AZD 8330); and 3,4-Difluoro-2-[(2-fluoro-4-iodophenyl)amino]-N-(2-hydroxyethoxy)-5-[(3-oxo-[1,2]oxazinan-2-yl)methyl]benzamide (CH 4987655 or Ro 4987655).

c-MET Inhibitors

In some embodiments, a c-MET inhibitor is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. c-MET, a receptor tyrosine kinase overexpressed or mutated in many tumor cell types, plays key roles in tumor cell proliferation, survival, invasion, metastasis, and tumor angiogenesis. Inhibition of c-MET may induce cell death in tumor cells overexpressing c-MET protein or expressing constitutively activated c-MET protein.

In some embodiments, the c-MET inhibitor is chosen from capmatinib (1NC280), JNJ-3887605, AMG 337, LY2801653, MSC2156119J, crizotinib, tivantinib, or golvatinib.

Exemplary c-MET Inhibitors

In some embodiments, the c-MET inhibitor comprises capmatinib (1NC280), or a compound described in U.S. Pat. Nos. 7,767,675, and 8,461,330, which are incorporated by reference in their entirety.

Other Exemplary c-MET Inhibitors

In some embodiments, the c-MET inhibitor comprises JNJ-38877605. JNJ-38877605 is an orally available, small molecule inhibitor of c-Met. JNJ-38877605 selectively binds to c-MET, thereby inhibiting c-MET phosphorylation and disrupting c-Met signal transduction pathways.

In some embodiments, the c-Met inhibitor is AMG 208. AMG 208 is a selective small-molecule inhibitor of c-MET. AMG 208 inhibits the ligand-dependent and ligand-independent activation of c-MET, inhibiting its tyrosine kinase activity, which may result in cell growth inhibition in tumors that overexpress c-Met.

In some embodiments, the c-Met inhibitor comprises AMG 337. AMG 337 is an orally bioavailable inhibitor of c-Met. AMG 337 selectively binds to c-MET, thereby disrupting c-MET signal transduction pathways.

In some embodiments, the c-Met inhibitor comprises LY2801653. LY2801653 is an orally available, small molecule inhibitor of c-Met. LY2801653 selectively binds to c-MET, thereby inhibiting c-MET phosphorylation and disrupting c-Met signal transduction pathways.

In some embodiments, c-Met inhibitor comprises MSC2156119J. MSC2156119J is an orally bioavailable inhibitor of c-Met. MSC2156119J selectively binds to c-MET, which inhibits c-MET phosphorylation and disrupts c-Met-mediated signal transduction pathways.

In some embodiments, the c-MET inhibitor is capmatinib. Capmatinib is also known as INCB028060. Capmatinib is an orally bioavailable inhibitor of c-MET. Capmatinib selectively binds to c-Met, thereby inhibiting c-Met phosphorylation and disrupting c-Met signal transduction pathways.

In some embodiments, the c-MET inhibitor comprises crizotinib. Crizotinib is also known as PF-02341066. Crizotinib is an orally available aminopyridine-based inhibitor of the receptor tyrosine kinase anaplastic lymphoma kinase (ALK) and the c-Met/hepatocyte growth factor receptor (HGFR). Crizotinib, in an ATP-competitive manner, binds to and inhibits ALK kinase and ALK fusion proteins. In addition, crizotinib inhibits c-Met kinase, and disrupts the c-Met signalling pathway. Altogether, this agent inhibits tumor cell growth.

In some embodiments, the c-MET inhibitor comprises golvatinib. Golvatinib is an orally bioavailable dual kinase inhibitor of c-MET and VEGFR-2 with potential antineoplastic activity. Golvatinib binds to and inhibits the activities of both c-MET and VEGFR-2, which may inhibit tumor cell growth and survival of tumor cells that overexpress these receptor tyrosine kinases.

In some embodiments, the c-MET inhibitor is tivantinib. Tivantinib is also known as ARQ 197. Tivantinib is an orally bioavailable small molecule inhibitor of c-MET. Tivantinib binds to the c-MET protein and disrupts c-Met signal transduction pathways, which may induce cell death in tumor cells overexpressing c-MET protein or expressing constitutively activated c-Met protein.

TGF-β Inhibitors

In some embodiments, a transforming growth factor beta (also known as TGF-β TGFβ, TGFb, or TGF-beta, used interchangeably herein) inhibitor is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In certain embodiments, a combination described herein comprises a transforming growth factor beta (also known as TGF-β TGFβ, TGFb, or TGF-beta, used interchangeably herein) inhibitor.

TGF-β belongs to a large family of structurally-related cytokines including, e.g., bone morphogenetic proteins (BMPs), growth and differentiation factors, activins and inhibins. In some embodiments, the TGF-3 inhibitors described herein can bind and/or inhibit one or more isoforms of TGF-β (e.g., one, two, or all of TGF-β1, TGF-β2, or TGF-β3).

Under normal conditions, TGF-β maintains homeostasis and limits the growth of epithelial, endothelial, neuronal and hematopoietic cell lineages, e.g., through the induction of anti-proliferative and apoptotic responses. Canonical and non-canonical signalling pathways are involved in cellular responses to TGF-β. Activation of the TGF-β/Smad canonical pathway can mediate the anti-proliferative effects of TGF-β. The non-canonical TGF-β pathway can activate additional intra-cellular pathways, e.g., mitogen-activated protein kinases (MAPK), phosphatidylinositol 3 kinase/Protein Kinase B, Rho-like GTPases (Tian et al. Cell Signal. 2011; 23(6):951-62; Blobe et al. N Engl J Med. 2000; 342(18):1350-8), thus modulating epithelial to mesenchymal transition (EMT) and/or cell motility.

Alterations of TGF-β signalling pathway are associated with human diseases, e.g., cancers, cardio-vascular diseases, fibrosis, reproductive disorders, and wound healing. Without wishing to be bound by theory, it is believed that in some embodiments, the role of TGF-β in cancer is dependent on the disease setting (e.g., tumor stage and genetic alteration) and/or cellular context. For example, in late stages of cancer, TGF-β can modulate a cancer-related process, e.g., by promoting tumor growth (e.g., inducing EMT), blocking anti-tumor immune responses, increasing tumor-associated fibrosis, or enhancing angiogenesis (Wakefield and Hill Nat Rev Cancer. 2013; 13(5):328-41). In certain embodiments, a combination comprising a TGF-β inhibitor described herein is used to treat a cancer in a late stage, a metastatic cancer, or an advanced cancer.

Preclinical evidence indicates that TGF-β plays an important role in immune regulation (Wojtowicz-Praga Invest New Drugs. 2003; 21(1):21-32; Yang et al. Trends Immunol. 2010; 31(6):220-7). TGF-β can down-regulate the host-immune response via several mechanisms, e.g., shift of the T-helper balance toward Th2 immune phenotype; inhibition of anti-tumoral Th1 type response and M1-type macrophages; suppression of cytotoxic CD8+T lymphocytes (CTL), NK lymphocytes and dendritic cell functions, generation of CD4+ CD25+T-regulatory cells; or promotion of M2-type macrophages with pro-tumoral activity mediated by secretion of immunosuppressive cytokines (e.g., IL10 or VEGF), pro-inflammatory cytokines (e.g., IL6, TNFα, or IL1) and generation of reactive oxygen species (ROS) with genotoxic activity (Yang et al. Trends Immunol. 2010; 31(6):220-7; Truty and Urrutia Pancreatology. 2007; 7(5-6):423-35; Achyut et al Gastroenterology. 2011; 141(4):1167-78).

Exemplary TGF-β Inhibitors

In some embodiments, the TGF-β inhibitor comprises XOMA 089, or a compound disclosed in International Application Publication No. WO 2012/167143, which is incorporated by reference in its entirety.

XOMA 089 is also known as XPA.42.089. XOMA 089 is a fully human monoclonal antibody that specifically binds and neutralizes TGF-beta 1 and 2 ligands.

The heavy chain variable region of XOMA 089 has the amino acid sequence of: QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKF QGRVTITADESTSTAYMELSSLRSEDTAVYYCARGLWEVRALPSVYWGQGTLVTVSS (SEQ ID NO: 284) (disclosed as SEQ ID NO: 6 in WO 2012/167143). The light chain variable region of XOMA 089 has the amino acid sequence of: SYELTQPPSVSVAPGQTARITCGANDIGSKSVHWYQQKAGQAPVLVVSEDIIRPSGIPERISGSNSG NTATLTISRVEAGDEADYYCQVWDRDSDQYVFGTGTKVTVLG (SEQ ID NO: 285) (disclosed as SEQ ID NO: 8 in WO 2012/167143).

XOMA 089 binds with high affinity to the human TGF-β isoforms. Generally, XOMA 089 binds with high affinity to TGF-β1 and TGF-β2, and to a lesser extent to TGF-β3. In Biacore assays, the K_(D) of XOMA 089 on human TGF-β is 14.6 pM for TGF-β1, 67.3 pM for TGF-β2, and 948 pM for TGF-33. Given the high affinity binding to all three TGF-β isoforms, in certain embodiments, XOMA 089 is expected to bind to TGF-β1, 2 and 3 at a dose of XOMA 089 as described herein. XOMA 089 cross-reacts with rodent and cynomolgus monkey TGF-β and shows functional activity in vitro and in vivo, making rodent and cynomolgus monkey relevant species for toxicology studies.

Other Exemplary TGF-β Inhibitors

In some embodiments, the TGF-β inhibitor comprises fresolimumab (CAS Registry Number: 948564-73-6). Fresolimumab is also known as GC1008. Fresolimumab is a human monoclonal antibody that binds to and inhibits TGF-beta isoforms 1, 2 and 3.

The heavy chain of fresolimumab has the amino acid sequence of:

(SEQ ID NO: 280) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQGLEWMGG VIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRSEDTAVYYCASTL GLVLDAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVK DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKT YTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK.

The light chain of fresolimumab has the amino acid sequence of:

(SEQ ID NO: 281) ETVLTQSPGTLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQAPRLLIY GASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYADSPITFG QGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC.

Fresolimumab is disclosed, e.g., in International Application Publication No. WO 2006/086469, and U.S. Pat. Nos. 8,383,780 and 8,591,901, which are incorporated by reference in their entirety.

IL-1β Inhibitors

The Interleukin-1 (IL-1) family of cytokines is a group of secreted pleotropic cytokines with a central role in inflammation and immune response. Increases in IL-1 are observed in multiple clinical settings including cancer (Apte et al. (2006) Cancer Metastasis Rev. p. 387-408; Dinarello (2010) Eur. J. Immunol. p. 599-606). The IL-1 family comprises, inter alia, IL-1 beta (IL-1b), and IL-1alpha (IL-1a). IL-1b is elevated in lung, breast and colorectal cancer (Voronov et al. (2014) Front Physiol. p. 114) and is associated with poor prognosis (Apte et al. (2000) Adv. Exp. Med. Biol. p. 277-88). Without wishing to be bound by theory, it is believed that in some embodiments, secreted IL-1b, derived from the tumor microenvironment and by malignant cells, promotes tumor cell proliferation, increases invasiveness and dampens anti-tumor immune response, in part by recruiting inhibitory neutrophils (Apte et al. (2006) Cancer Metastasis Rev. p. 387-408; Miller et al. (2007) J. Immunol. p. 6933-42). Experimental data indicate that inhibition of IL-1b results in a decrease in tumor burden and metastasis (Voronov et al. (2003) Proc. Natl. Acad. Sci. U.S.A. p. 2645-50).

In some embodiments, an interleukin-1 beta (IL-1(3) inhibitor is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. In some embodiments, the IL-13 inhibitor is chosen from canakinumab, gevokizumab, Anakinra, or Rilonacept. In some embodiments, the IL-1β inhibitor is canakinumab.

Exemplary IL-1 Inhibitors

In some embodiments, the IL-1(3 inhibitor is canakinumab. Canakinumab is also known as ACZ885 or ILARIS®. Canakinumab is a human monoclonal IgG1/K antibody that neutralizes the bioactivity of human IL-1β.

Canakinumab is disclosed, e.g., in WO 2002/16436, U.S. Pat. No. 7,446,175, and EP 1313769. The heavy chain variable region of canakinumab has the amino acid sequence of: MEFGLSWVFLVALLRGVQCQVQLVESGGGVVQPGRSLRLSCAASGFTFSVYGMNWVRQAPGK GLEWVAIIWYDGDNQYYADSVKGRFTISRDNSKNTLYLQMNGLRAEDTAVYYCARDLRTGPFD YWGQGTLVTVSS (SEQ ID NO: 282) (disclosed as SEQ ID NO: 1 in U.S. Pat. No. 7,446,175). The light chain variable region of canakinumab has the amino acid sequence of: MLPSQLIGFLLLWVPASRGEIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLI KYASQSFSGVPSRFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPFTFGPGTKVDIK (SEQ ID NO: 283) (disclosed as SEQ ID NO: 2 in U.S. Pat. No. 7,446,175).

Canakinumab has been used, e.g., for the treatment of Cryopyrin Associated Periodic Syndromes (CAPS), in adults and children, for the treatment of systemic juvenile idiopathic arthritis (SJIA), for the symptomatic treatment of acute gouty arthritis attacks in adults, and for other IL-1β driven inflammatory diseases. Without wishing to be bound by theory, it is believed that in some embodiments, IL-1β inhibitors, e.g., canakinumab, can increase anti-tumor immune response, e.g., by blocking one or more functions of IL-1b including, e.g., recruitment of immunosuppressive neutrophils to the tumor microenvironment, stimulation of tumor angiogenesis, and/or promotion of metastasis (Dinarello (2010) Eur. J. Immunol. p. 599-606).

In some embodiments, the combination described herein includes an IL-1(3 inhibitor, canakinumab, or a compound disclosed in WO 2002/16436, and an inhibitor of an immune checkpoint molecule, e.g., an inhibitor of PD-1 (e.g., an anti-PD-1 antibody molecule). IL-1 is a secreted pleotropic cytokine with a central role in inflammation and immune response. Increases in IL-1 are observed in multiple clinical settings including cancer (Apte et al. (2006) Cancer Metastasis Rev. p. 387-408; Dinarello (2010) Eur. J. Immunol. p. 599-606). IL-1b is elevated in lung, breast and colorectal cancer (Voronov et al. (2014) Front Physiol. p. 114) and is associated with poor prognosis (Apte et al. (2000) Adv. Exp. Med. Biol. p. 277-88). Without wishing to be bound by theory, it is believed that in some embodiments, secreted IL-1b, derived from the tumor microenvironment and by malignant cells, promotes tumor cell proliferation, increases invasiveness and dampens anti-tumor immune response, in part by recruiting inhibitory neutrophils (Apte et al. (2006) Cancer Metastasis Rev. p. 387-408; Miller et al. (2007) J. Immunol. p. 6933-42). Experimental data indicate that inhibition of IL-1b results in a decrease in tumor burden and metastasis (Voronov et al. (2003) Proc. Natl. Acad. Sci. U.S.A. p. 2645-50). Canakinumab can bind IL-1b and inhibit IL-1-mediated signalling. Accordingly, in certain embodiments, an IL-1β inhibitor, e.g., canakinumab, enhances, or is used to enhance, an immune-mediated anti-tumor effect of an inhibitor of PD-1 (e.g., an anti-PD-1 antibody molecule).

In some embodiments, the IL-1β inhibitor, canakinumab, or a compound disclosed in WO 2002/16436, and the inhibitor of an immune checkpoint molecule, e.g., an inhibitor of PD-1 (e.g., an anti-PD-1 antibody molecule), each is administered at a dose and/or on a time schedule, that in combination, achieves a desired anti-tumor activity.

MDM2 Inhibitors

In some embodiments, a mouse double minute 2 homolog (MDM2) inhibitor is used in combination with the compounds of Formula (I′) or Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof, for treating a disease, e.g., cancer. The human homolog of MDM2 is also known as HDM2. In some embodiments, an MDM2 inhibitor described herein is also known as a HDM2 inhibitor. In some embodiments, the MDM2 inhibitor is chosen from HDM201 or CGM097.

In an embodiment the MDM2 inhibitor comprises (S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one (also known as CGM097) or a compound disclosed in PCT Publication No. WO 2011/076786 to treat a disorder, e.g., a disorder described herein). In one embodiment, a therapeutic agent disclosed herein is used in combination with CGM097.

In an embodiment, an MDM2 inhibitor comprises an inhibitor of p53 and/or a p53/Mdm2 interaction. In an embodiment, the MDM2 inhibitor comprises (S)-5-(5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4-chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol-4(1H)-one (also known as HDM201), or a compound disclosed in PCT Publication No. WO2013/111105 to treat a disorder, e.g., a disorder described herein. In one embodiment, a therapeutic agent disclosed herein is used in combination with HDM201. In some embodiments, HDM201 is administered orally.

In one embodiment, the combination disclosed herein is suitable for the treatment of cancer in vivo. For example, the combination can be used to inhibit the growth of cancerous tumors. The combination can also be used in combination with one or more of: a standard of care treatment (e.g., for cancers or infectious disorders), a vaccine (e.g., a therapeutic cancer vaccine), a cell therapy, a radiation therapy, surgery, or any other therapeutic agent or modality, to treat a disorder herein. For example, to achieve antigen-specific enhancement of immunity, the combination can be administered together with an antigen of interest.

EXAMPLES

The disclosure is further illustrated by the following examples and synthesis schemes, which are not to be construed as limiting this disclosure in scope or spirit to the specific procedures herein described. It is to be understood that the examples are provided to illustrate certain embodiments and that no limitation to the scope of the disclosure is intended thereby. It is to be further understood that resort may be had to various other embodiments, modifications, and equivalents thereof which may suggest themselves to those skilled in the art without departing from the spirit of the present disclosure and/or scope of the appended claims.

Compounds of the present disclosure may be prepared by methods known in the art of organic synthesis. In all of the methods it is understood that protecting groups for sensitive or reactive groups may be employed where necessary in accordance with general principles of chemistry. Protecting groups are manipulated according to standard methods of organic synthesis (T. W. Green and P. G. M. Wuts (1999) Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons). These groups are removed at a convenient stage of the compound synthesis using methods that are readily apparent to those skilled in the art.

Analytical Methods, Materials, and Instrumentation

Unless otherwise noted, reagents and solvents were used as received from commercial suppliers. Proton nuclear magnetic resonance (NMR) spectra were obtained on either Bruker Avance spectrometer or Varian Oxford 400 MHz spectrometer unless otherwise noted. Spectra are given in ppm (δ) and coupling constants, J, are reported in Hertz. Tetramethylsilane ((CH₃)₄Si) was used as an internal standard. Chemical shifts are reported in ppm relative to dimethyl sulfoxide (δ 2.50), methanol (δ 3.31), chloroform (δ 7.26) or other solvent as indicated in NMR spectral data. A small amount of the dry sample (2-5 mg) is dissolved in an appropriate deuterated solvent (1 mL). The chemical names were generated using ChemBioDraw Ultra v14 from CambridgeSoft.

Mass spectra (ESI-MS) were collected using a Waters System (Acquity UPLC and a Micromass ZQ mass spectrometer) or Agilent-1260 Infinity (6120 Quadrupole); all masses reported are the m/z of the protonated parent ions unless recorded otherwise. The sample was dissolved in acquirable solvent such as MeCN, DMSO, or MeOH and was injected directly into the column using an automated sample handler. Abbreviations used in the following examples and elsewhere herein are:

-   -   AgOTf silver trifluoromethanesulfonate     -   AIBN azobisisobutyronitrile     -   aq aqueous     -   Boc tert-butyloxycarbonyl     -   Bu butyl     -   Bu₄NI or TBAI tetrabutylammonium iodide     -   br broad     -   DAST diethylaminosulfur trifluoride     -   DBAD di-tert-butyl azodicarboxylate     -   DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene     -   DCM dichloromethane or methylene chloride     -   d doublet     -   dd doublet of doublets     -   ddd doublet of doublet of doublets     -   dddd doublet of doublet of doublets of doublets     -   DHP dihydropyran     -   DIAD diisopropyl azodicarboxylate     -   DMA N,N-dimethylacetamide     -   DMAP 4-Dimethylaminopyridine     -   DME 1,2-dimethoxyethane (glyme)     -   DMF N,N-dimethylformamide     -   DMP Dess-Martin periodinane     -   DMSO dimethylsulfoxide     -   dq doublet of quartets     -   dt doublet of triplets     -   dtd doublet of triplet of doublets     -   dtbbpy 4,4′-di-tert-butyl-2,2′-bipyridine     -   EC₅₀ half maximal effective concentration (qualified)     -   Et ethyl     -   Et₂O diethyl ether     -   EtOAc ethyl acetate     -   EtOH ethanol     -   Et₃SiH triethylsilane     -   g gram     -   h or hr hour     -   hept heptet     -   heptd heptet of doublets     -   HPLC high performance liquid chromatography     -   HRMS high resonance mass spectrometry     -   i-Pr isopropyl     -   i-Pr₂NEt or DIPEA N,N-diisopropylethylamine     -   IPA Iso-propanol     -   Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆         4,4′-Bis(1,1-dimethylethyl)-2,2′-bipyridine-N1,N1′]bis[3,5-difluoro-2-[5-(trifluoromethyl)-2-pyridinyl-N]phenyl-C]Iridium(III)         hexafluorophosphate     -   LCMS liquid chromatography mass spectrometry     -   LDA lithium diisopropylamide     -   LED light-emitting diode     -   M molar     -   m multiplet     -   Me methyl     -   MeCN acetonitrile     -   MeOH methanol     -   mg milligram     -   MHz megahertz     -   min minutes     -   mL milliliter     -   mmol millimole     -   Ms methanesulfonyl     -   MsOH methanesulfonic acid     -   MS mass spectrometry     -   MTBE methyl tert-butyl ether     -   μW microwave     -   NaBH(OAc)₃ sodium triacetoxyborohydride     -   NaHCO₃ sodium carbonate     -   Na₂SO₄ sodium sulfate     -   NBS N-bromosuccinimide     -   NMP 1-methyl-2-pyrrolidinone     -   NMR Nuclear magnetic resonance     -   O/N overnight     -   Ph phenyl     -   PhMe toluene     -   Pyr pyridine     -   q quartet     -   qd quartet of doublets     -   quint quintet     -   quintd quintet of doublets     -   rt room temperature     -   Rt retention time     -   RuPhos 2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl     -   s singlet     -   sat. saturated     -   SFC supercritical fluid chromatography     -   SPA scintillation proximity assay     -   SOCl₂ thionyl chloride     -   SEM 2-(trimethylsilyl)ethoxymethyl     -   t triplet     -   t-Bu or tBu tert-butyl     -   TBAB tetrabutylamonium bromide     -   TBAF tetrabutylamonium fluoride     -   TBS tert-butyldimethylsilyl     -   t-BuONa sodium tert-butoxidetd triplet of doublets     -   tdd triplet of doublet of doublets     -   TEA or Et₃N or NEt₃ triethyl amine     -   Tf trifluoromethanesulfonate or triflate     -   THF tetrahydrofuran     -   TFA or CF₃CO₂H trifluoroacetic acid     -   TFE 2,2,2-trifluoroethanol     -   Ti(i-OPr)₄ titanium(IV) isopropoxide     -   TLC thin layer chromatography     -   TMP 2,2,6,6-tetramethylpiperidine     -   TMS trimethylsilyl     -   Ts tosyl     -   ttd triplet of triplet of doublets     -   UPLC Ultra-Performance Liquid Chromatography

Example 1: 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HCl or CF₃CO₂H Salt (I-12)

Step 1. Ethyl 4-bromo-2-(chloromethyl)benzoate (I-1b)

A stirred suspension of 5-bromophthalide (1-1a, 1200 g, 5.633 mol) in EtOH (12 L) was heated to 68-72° C. SOCl₂ (2.40 L, 33.0 mol) was then added dropwise over a period of 7 h. The reaction mixture was concentrated under reduced pressure to about 4 L, and then water (5 L) and MTBE (5 L) were added. The resulting mixture was stirred for 40 min. The phases were separated and the aqueous phase was extracted with MTBE (1×5 L). The combined organic layers were washed with 5% aq. NaHCO₃ (5 L), dried over Na₂SO₄, filtered, and concentrated to dryness to afford 1-1b (1450 g, 5.25 mol, 93% yield) as a pale brown solid. MS [M+Na]⁺=298.9. ¹H NMR (400 MHz, Chloroform-d) δ 7.85 (d, J=8.4 Hz, 1H), 7.72 (d, J=2.0 Hz, 1H), 7.52 (dd, J=8.3, 2.0 Hz, 1H), 5.00 (s, 2H), 4.38 (q, J=7.1 Hz, 2H), 1.40 (t, J=7.1 Hz, 3H).

Step 2. 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (1-1d)

To a stirred suspension of 3-aminopiperidine-2,6-dione hydrochloride (1-1c, 596.3 g, 3.623 mol) and i-Pr₂NEt (2.50 L, 14.3 mol) in DMF (5.0 L) was added 1-1b (1000 g, 3.623 mmol) and the resulting reaction mixture was stirred at 85-90° C. for 24 h. The reaction mixture was then allowed to cool to room temperature, water (20 L) was added, and the resulting mixture was stirred for 12 h. The formed precipitate was filtered and washed with water (5 L) and MeOH (2 L). The crude solid was slurried in MeOH (5 L) for 1 h, filtered, and washed with MeOH (2 L). The resulting solid was then taken in EtOAc (10 L) and stirred for 1 h. The obtained suspension was then filtered, washed with EtOAc (5 L), and dried under reduced pressure at 45-50° C. to afford 1-1d (740 g, 2.29 mol, 63% yield) as an off-white solid. MS [M+1]=323.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.99 (s, 1H), 7.91-7.88 (m, 1H), 7.72 (dd, J=8.1, 1.6 Hz, 1H), 7.67 (d, J=8.0 Hz, 1H), 5.11 (dd, J=13.3, 5.1 Hz, 1H), 4.47 (d, J=17.7 Hz, 1H), 4.34 (d, J=17.7 Hz, 1H), 2.98-2.83 (m, 1H), 2.65-2.55 (m, 1H), 2.45-2.29 (m, 1H), 2.01 (dtd, J=12.7, 5.3, 2.3 Hz, 1H).

Step 3. tert-butyl ((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)carbamate (1-1f)

To a stirred suspension of 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (1-1d, 2.65 g, 8.20 mmol), tert-butyl ((1S,2S)-2-hydroxycyclopentyl)carbamate (1-1e, 1.50 g, 7.45 mmol), NiCl₂(glyme) (0.082 g, 0.37 mmol), dtbbpy (0.100 g, 0.373 mmol) and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (0.083 g, 0.075 mmol) in MeCN (25 mL) under a nitrogen atmosphere was added 2,2,6,6-tetramethylpiperidine (40-2, 1.90 mL, 11.2 mmol). The resulting mixture was then stirred vigorously for 70 h under irradiation of blue LED light at room temperature. The reaction mixture was then diluted with DCM (20 mL), filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 12% EtOH:EtOAc (v/v=1:3) in DCM to afford 1-1f (1.03 g, 2.11 mmol, 28% yield) as a white powder. MS [M-tBu+H]⁺=388.1. ¹H NMR (400 MHz, DCM-d₂) δ 8.02 (s, 1H), 7.68 (d, J=8.4 Hz, 1H), 7.15 (s, 1H), 7.05 (d, J=8.5 Hz, 1H), 5.11 (ddd, J=13.1, 5.1, 2.8 Hz, 1H), 4.68-4.61 (m, 1H), 4.57 (s, 1H), 4.40-4.25 (m, 2H), 4.04-3.97 (m, 1H), 2.95-2.71 (m, 2H), 2.41-2.24 (m, 1H), 2.21-1.98 (m, 3H), 1.89-1.70 (m, 3H), 1.53-1.44 (m, 1H), 1.41 (s, 9H).

Step 4. Method A. 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (I-12)

To a stirred solution of tert-butyl ((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)carbamate (1-1f, 1.50 g, 2.99 mmol) in THF (5 mL) was added 4 M HCl in dioxane (2.0 mL, 8.0 mmol). Obtained reaction mixture was stirred for 2 h at 60° C. Formation of white precipitate was observed. The reaction mixture was diluted with Et₂O (15 mL) and filtered. The precipitate was washed with Et₂O (4×) and then dried under reduced pressure to afford the hydrochloride salt of I-12 (1.01 g, 2.66 mmol, 89% yield) as a white solid. MS [M+H]⁺=344.1. ¹H NMR (400 MHz, DMSO-d₆) δ 10.98 (s, 1H), 8.36 (s, 2H), 7.68-7.63 (m, 1H), 7.34-7.18 (m, 1H), 7.08 (dt, J=8.4, 2.3 Hz, 1H), 5.08 (dd, J=13.3, 5.1 Hz, 1H), 4.95-4.82 (m, 1H), 4.41 (dd, J=17.3, 3.5 Hz, 1H), 4.28 (dd, J=17.3, 2.8 Hz, 1H), 3.62 (s, 1H), 2.99-2.83 (m, 1H), 2.69-2.54 (m, 1H), 2.47-2.31 (m, 1H), 2.31-2.21 (m, 1H), 2.18-2.06 (m, 1H), 2.04-1.94 (m, 1H), 1.89-1.62 (m, 4H).

Step 5. Method B. 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione CF₃CO₂H Salt (I-12)

To a stirred solution of tert-butyl ((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)carbamate (1-1f, 573 mg, 1.29 mmol) in DCM (4.3 mL) was added CF₃CO₂H (1.1 mL, 13 mmol) and the resulting mixture was stirred at room temperature for 5 h. The solution was concentrated under reduced pressure and azeotroped with PhMe to afford the desired crude product (TFA salt of I-12) as a light yellow foam (assumed quantitative yield). The crude product carried onto the next step without further purification. MS [M+H]⁺=344.4. ¹H NMR (400 MHz, Methanol-d₄) δ 7.74 (dd, J=8.4, 1.3 Hz, 1H), 7.16 (d, J=0.7 Hz, 1H), 7.13-7.09 (m, 1H), 5.12 (ddd, J=13.4, 5.2, 2.7 Hz, 1H), 4.86-4.76 (m, 1H), 4.50-4.37 (m, 2H), 3.77 (td, J=8.1, 5.3 Hz, 1H), 2.90 (ddd, J=17.6, 13.5, 5.3 Hz, 1H), 2.78 (ddd, J=17.6, 4.7, 2.4 Hz, 1H), 2.55-2.40 (m, 1H), 2.42-2.23 (m, 2H), 2.16 (dtd, J=12.9, 5.3, 2.4 Hz, 1H), 1.99-1.87 (m, 2H), 1.87-1.74 (m, 1H), 1.79-1.65 (m, 1H).

Alternatively, Conversion of 1-1d to I-12 was Also Achieved Via the Following Synthetic Procedure:

Step 1. 3-(5-bromo-1-oxoisoindolin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (31-3a)

To a stirred solution of 1-1d (10.0 g, 30.9 mmol) and DBU (6.9 mL, 46 mmol) in DMF (95 mL) was added SEMCl (6.6 mL, 37 mmol) at 0° C. and the resulting reaction mixture was allowed to warm to room temperature and then stirred for 5 h. An additional portion of DBU (3.5 mL, 23 mmol) and SEMCl (3.3 mL, 19 mmol) was added and stirring was continued for an additional 2 h. The reaction mixture was then quenched with sat. aq. NH₄Cl (250 mL) and extracted with EtOAc (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was dissolved in minimal amount of EtOAc (˜50 mL) and Et₂O:heptane (v/v=1:2, 400 mL) was added. The resulting cloudy solution was left standing at −5° C. overnight. The formed precipitate was filtered, washed with heptane (×3), and dried under vacuum to afford 31-3a (11.53 g, 25.4 mmol, 82% yield) as an off-white solid. MS [M+H]⁺=453.4. ¹H NMR (400 MHz, Chloroform-d) δ 7.75 (d, J=8.6 Hz, 1H), 7.66-7.61 (m, 2H), 5.37-5.09 (m, 3H), 4.48 (d, J=16.2 Hz, 1H), 4.32 (d, J=16.2 Hz, 1H), 3.74-3.50 (m, 2H), 3.11-2.98 (m, 1H), 2.94-2.83 (m, 1H), 2.33 (qd, J=13.2, 4.7 Hz, 1H), 2.24-2.15 (m, 1H), 0.97-0.90 (m, 2H), 0.00 (s, 9H).

Step 2. tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)carbamate (1-1g)

To a stirred solution of 31-3a (2.00 g, 4.41 mmol), 1-1e (0.888 g, 4.41 mmol), NiCl₂(glyme) (0.048 g, 0.22 mmol), dtbbpy (0.059 g, 0.22 mmol) and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (0.049 g, 0.044 mmol) in degassed MeCN (16 mL) under an atmosphere of nitrogen was added 2,2,6,6-tetramethylpiperidine (40-2, 0.75 mL, 4.4 mmol) and the resulting mixture was stirred vigorously for 24 h under irradiation of blue LED lights at room temperature. The reaction mixture was then filtered and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford 1-Ig (2.19 g, 3.82 mmol, 87% yield) as a yellow solid. MS [M−H]⁻=572.5. ¹H NMR (400 MHz, DMSO-d₆) δ 7.63 (d, J=8.4 Hz, 1H), 7.22 (s, 1H), 7.11-7.05 (m, 1H), 5.29-5.16 (m, 1H), 5.12-5.02 (m, 2H), 4.69-4.64 (m, 1H), 4.50-4.36 (m, 1H), 4.24 (dd, J=17.0, 9.1 Hz, 1H), 3.95-3.85 (m, 1H), 3.64-3.47 (m, 2H), 3.13-3.01 (m, 1H), 2.87-2.74 (m, 1H), 2.49-2.31 (m, 1H), 2.12-1.97 (m, 3H), 1.86-1.60 (m, 3H), 1.59-1.47 (m, 1H), 1.40 (s, 9H), 0.90-0.83 (m, 2H), 0.00 (s, 9H).

Step 3. 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12)

To a solution of 1-1g (6.85 g, 9.55 mmol) in DCM (53 mL) under an atmosphere of nitrogen was added methanesulfonic acid (2.5 mL, 38 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was cooled to 0° C. and triethylamine (10.7 mL, 76 mmol) and N1,N2-dimethylethane-1,2-diamine (1.23 mL, 11.5 mmol) were added and stirring was continued at rt for 4 h. Additional N1,N2-dimethylethane-1,2-diamine (0.51 mL, 4.8 mmol) was added and stirring was continued at rt for 30 minutes. The reaction mixture was quenched with a 50% saturated aqueous sodium hydrogen carbonate and extracted with DCM:isopropanol (v/v=4:1, ×4). The organic phases were combined, passed through a phase separator and concentrated to dryness. The material was suspended in minimal DCM and excess diethyl ether was added. The resulting suspension was sonicated for 1 hr, filtered, and rinsed with diethyl ether (×3). The solid was collected and dried in a high vacuum oven at 50° C. overnight to afford I-12 (3.02 g, 8.80 mmol, 92% yield) as a cream solid. MS [M+H]⁺=344.1. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.19-7.14 (m, 1H), 7.03 (dd, J=8.2, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.47-4.33 (m, 2H), 4.25 (d, J=17.1 Hz, 1H), 3.34-3.25 (m, 3H), 2.91 (ddd, J=17.5, 13.5, 5.3 Hz, 1H), 2.64-2.54 (m, 1H), 2.37 (qd, J=13.1, 4.1 Hz, 1H), 2.26-2.11 (m, 1H), 2.03-1.84 (m, 2H), 1.82-1.58 (m, 3H), 1.42-1.32 (m, 1H).

Example 2: 3-(5-(((1S,2S)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-13)

To a stirred solution of the HCl salt of I-12 (500 mg, 1.24 mmol) and NaBH(OAc)₃ (787 mg, 3.71 mmol) in DMF (10 mL) was added acetaldehyde (2-1, 0.21 mL, 3.7 mmol) in one portion and the resulting mixture was stirred for 2 h at room temperature. The reaction mixture was then concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0% to 10% Et₃N in EtOAc to afford I-13 (342 mg, 0.85 mmol, 69% yield) as a white powder. MS [M+H]⁺=400.0. ¹H NMR (400 MHz, DCM-d₂) δ 8.66 (s, 1H), 7.69-7.65 (m, 1H), 7.14-6.97 (m, 2H), 5.10 (ddd, J=13.3, 5.2, 3.5 Hz, 1H), 4.71-4.52 (m, 1H), 4.39-4.18 (m, 2H), 3.40-3.23 (m, 1H), 2.89-2.72 (m, 2H), 2.66-2.47 (m, 4H), 2.41-2.19 (m, 1H), 2.20-2.07 (m, 1H), 2.02-1.87 (m, 2H), 1.83-1.62 (m, 3H), 1.58-1.45 (m, 1H), 1.05-0.94 (m, 6H).

Example 3: 3-(1-oxo-5-(((1S,2S)-2-(piperidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-32)

To a stirred solution of the HCl salt of I-12 (400 mg, 1.05 mmol) and NaBH(OAc)₃ (670 mg, 3.16 mmol) in DMF (10 mL) was added 25% glutaraldehyde in H₂O (0.42 mL, 1.1 mmol) and the resulting mixture was stirred for 40 min at room temperature. The reaction mixture was then concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0% to 10% Et₃N in EtOAc to afford I-32 (298 mg, 0.717 mmol, 68% yield) as a white powder. MS [M+H]⁺=412.1. ¹H NMR (400 MHz, DCM-d₂) δ 9.23 (br s, 1H), 7.84-7.52 (m, 1H), 7.08-6.81 (m, 2H), 5.18-4.94 (m, 1H), 4.81-4.60 (m, 1H), 4.32 (d, J=3.8 Hz, 1H), 4.14 (q, J=16.0 Hz, 1H), 3.10-2.71 (m, 3H), 2.57-2.25 (m, 4H), 2.20-2.05 (m, 2H), 2.04-1.88 (m, 2H), 1.79-1.62 (m, 3H), 1.60-1.46 (m, 5H), 1.45-1.34 (m, 2H).

Example 4: 3-(5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-29)

To a solution of the TFA salt of I-12 (19 mg, 0.055 mmol) in DCM-d₂ (0.5 mL) were added acetaldehyde (2-1, 0.02 mL, 0.3 mmol) and MgSO₄ (100 mg, 0.831 mmol) and the resulting mixture was stirred at room temperature for 5 h. The reaction mixture was then filtered and concentrated to dryness. The resulting material was taken up in DCM (0.5 mL) and NaBH(OAc)₃ (22 mg, 0.10 mmol) was added. After 30 min, the reaction was diluted with DCM and quenched with sat. aq. NaHCO₃. The phases were separated and the aqueous phase was extracted with EtOAc (5×5 mL). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 10 mM NH₄OH) and collected into tubes containing formic acid (3 drops). The fractions containing the desired product were combined and lyophilized to afford I-29 (8 mg, 0.018 mmol, 33% yield) as a white powder. MS [M+H]⁺=372.4. ¹H NMR (400 MHz, DCM-d₂) δ 7.61 (d, J=8.3 Hz, 1H), 6.93 (d, J=2.1 Hz, 1H), 6.91 (dd, J=8.3, 2.1 Hz, 1H), 5.02 (dt, J=13.3, 5.5 Hz, 1H), 4.76-4.70 (m, 1H), 4.31-4.15 (m, 2H), 3.33 (td, J=7.4, 4.0 Hz, 1H), 2.83-2.66 (m, 5H), 2.31-2.17 (m, 1H), 2.14-2.01 (m, 3H), 1.79-1.63 (m, 3H), 1.63-1.52 (m, 1H), 1.11 (t, J=7.1 Hz, 3H).

Example 5: 3-(5-(((1S,2S)-2-(3-hydroxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-5)

To a stirred solution of the TFA salt of I-12 (55 mg, 0.12 mmol) in MeCN (1 mL) was added i-Pr₂NEt (0.07 mL, 0.4 mmol) and epichlorohydrin (5-1a, 11 mg, 0.12 mmol) and the resulting reaction mixture was heated to 60° C. After 5 h the temperature was increased to 70° C. and the reaction mixture was stirred overnight. The mixture was then cooled to room temperature and another portion of epichlorohydrin (5-1a, 11 mg, 0.12 mmol) was added and the resulting mixture was heated to 70° C. for two days. The reaction mixture was cooled to room temperature and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 10 mM NH₄OH) and collected into tubes containing formic acid (3 drops). The fractions containing the desired product were combined and lyophilized to afford the formate salt of I-5 (11 mg, 0.023 mmol, 19.5% yield) as a white solid. MS [M+H]⁺=400.4. ¹H NMR (400 MHz, MeCN-d₃) δ 8.28 (s, 1H), 7.64 (d, J=8.4 Hz, 1H), 7.09-7.06 (m, 1H), 7.01 (dd, J=8.4, 2.3 Hz, 1H), 5.04 (ddd, J=13.4, 5.2, 2.0 Hz, 1H), 4.67 (dt, J=6.9, 4.0 Hz, 1H), 4.40-4.24 (m, 3H), 3.90 (dt, J=15.2, 7.6 Hz, 2H), 3.47-3.27 (m, 3H), 2.86-2.75 (m, 1H), 2.71 (ddd, J=17.6, 4.8, 2.6 Hz, 1H), 2.47-2.33 (m, 1H), 2.27-2.17 (m, 1H), 2.11 (dtd, J=13.0, 5.3, 2.6 Hz, 1H), 1.82-1.65 (m, 3H), 1.57-1.47 (m, 1H).

Example 6: 3-(5-(((1S,2S)-2-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-17)

Step 1. (1,3-dioxolane-2,2-diyl)bis(ethane-2,1-diyl) bis(4-methylbenzenesulfonate) (6-2a)

To as stirred solution of 2,2′-(1,3-dioxolane-2,2-diyl)diethanol (6-1a, 300 mg, 1.85 mmol) and Et₃N (0.65 mL, 4.6 mmol) in THF (6 mL) at −10° C. was added a solution of TsCl (882 mg, 4.62 mmol) in THF (1 mL) and the resulting mixture was allowed to warm to room temperature and then stirred for 6 days. The reaction mixture was then filtered and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 60% EtOAc in heptane to give 6-2a (426 mg, 0.905 mmol, 49% yield) as a white cloudy oil that crystallized upon standing. MS [M+H]⁺=471.3. ¹H NMR (400 MHz, Chloroform-d) δ 7.77 (d, J=8.3 Hz, 4H), 7.39-7.32 (m, 4H), 4.07 (t, J=6.9 Hz, 4H), 3.84 (s, 4H), 2.45 (s, 6H), 1.95 (t, J=6.9 Hz, 4H).

Step 2. 3-(5-(((1S,2S)-2-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-17)

To a stirred solution of the TFA salt of I-12 (25 mg, 0.055 mmol) and (1,3-dioxolane-2,2-diyl)bis(ethane-2,1-diyl) bis(4-methylbenzenesulfonate) (6-2a, 25 mg, 0.053 mmol) in DMF (1 mL) was added i-Pr₂NEt (0.05 mL, 0.3 mmol) and the resulting mixture was heated to 70° C. for two days. The reaction mixture was allowed to cool to room temperature and azeotroped with PhMe. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 10 mM NH₄OH) and collected into tubes containing formic acid (3 drops). The fractions containing the desired product were combined and lyophilized to afford the formate salt of I-17 (4 mg, 0.007 mmol, 13.5% yield) as a white powder. MS [M+H]⁺=470.5. ¹H NMR (400 MHz, MeCN-d₃) δ 8.22 (d, J=1.0 Hz, 1H), 7.64 (d, J=8.4 Hz, 1H), 7.13-7.09 (m, 1H), 7.01 (dd, J=8.5, 2.3 Hz, 1H), 5.03 (ddd, J=13.4, 5.2, 2.2 Hz, 1H), 4.70 (ddd, J=7.6, 4.6, 2.7 Hz, 1H), 4.36 (dd, J=16.8, 5.5 Hz, 1H), 4.28 (dd, J=16.8, 2.1 Hz, 1H), 3.87 (s, 4H), 3.03-2.96 (m, 1H), 2.86-2.75 (m, 1H), 2.71 (ddd, J=17.7, 4.8, 2.5 Hz, 1H), 2.63-2.55 (m, 4H), 2.45-2.40 (m, 1H), 2.14-2.05 (m, 2H), 1.74-1.68 (m, 3H), 1.65 (t, J=5.7 Hz, 4H), 1.54 (dtd, J=12.0, 9.4, 7.1 Hz, 1H).

Example 7: 3-(5-(((1S,2S)-2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-18)

Step 1. (cis-tetrahydrofuran-2,5-diyl)bis(methylene) bis(4-methylbenzenesulfonate) (7-2a)

To a stirred solution of cis-2,5-bishydroxymethyl-tetrahydrofuran (7-1a, 263 mg, 1.99 mmol) and DMAP (24 mg, 0.20 mmol) in THF (7 mL) was added Et₃N (1.3 mL, 9.3 mmol) and the resulting mixture was cooled to 0° C. Tosyl chloride (835 mg, 4.38 mmol) was added and the resulting mixture was allowed to warm to room temperature and then stirred overnight. The reaction mixture was then diluted with water (25 mL) and EtOAc (150 mL) was added. The organic layer was then washed with water (25 mL), sat. aq. NaHCO₃ (30 mL), and brine (30 mL), dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 60% EtOAc in heptane to afford the 7-2a as a white solid (488.6 mg, 1.098 mmol, 55% yield). MS [M+H₂O]⁺=458.4. ¹H NMR (400 MHz, DCM-d₂) δ 7.87-7.73 (m, 4H), 7.48-7.37 (m, 4H), 4.10 (dddd, J=11.1, 6.4, 4.3, 1.9 Hz, 2H), 4.02-3.85 (m, 4H), 2.49 (s, 6H), 2.07-1.91 (m, 2H), 1.76-1.61 (m, 2H).

Step 2. 3-(5-(((1S,2S)-2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-18)

To a stirred solution of the TFA salt of I-12 (54.9 mg, 0.120 mmol) and (cis-tetrahydrofuran-2,5-diyl)bis(methylene) bis(4-methylbenzenesulfonate) (7-2a, 68.3 mg, 0.155 mmol) in DMF (2 mL) was added i-Pr₂NEt (0.10 mL, 0.57 mmol) and the resulting mixture was stirred at room temperature for 2 days. The reaction mixture was then heated to 85° C. and stirred overnight. The resulting mixture was diluted with EtOAc (80 mL) and washed with sat. aq. NaHCO₃ (2×20 mL) and brine (20 mL). The organic layer was then dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The pure fractions were combined and lyophilized to afford the formate salt of I-18 as a white solid (2.3 mg, 4.5 μmol, 4% yield). MS [M+H]+=440.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.20 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.21 (t, J=2.7 Hz, 1H), 7.05 (dt, J=8.4, 1.9 Hz, 1H), 5.15-5.01 (m, 1H), 4.69 (d, J=6.9 Hz, 1H), 4.48-4.10 (m, 4H), 2.97-2.75 (m, 1H), 2.63-2.58 (m, 1H), 2.38 (dd, J=13.0, 4.3 Hz, 1H), 2.35-2.24 (m, 3H), 2.10-2.03 (m, 1H), 2.02-1.93 (m, 1H), 1.81 (d, J=7.4 Hz, 3H), 1.74-1.57 (m, 5H), 1.49 (d, J=12.2 Hz, 1H).

Example 8: 3-(5-(((1S,2S)-2-(ethyl(isopropyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-25)

To a stirred suspension of the HCl salt of I-12 (50 mg, 0.13 mmol), acetone (0.03 mL, 0.4 mmol) and MgSO₄ (32 mg, 0.26 mmol) in DMA (1.0 mL) was added NaBH(OAc)₃ (84 mg, 0.40 mmol) and the resulting mixture was stirred overnight at room temperature. Acetaldehyde (2-1, 0.04 mL, 0.7 mmol) was then added and the resulting mixture was stirred for an additional 4 h. The reaction mixture was diluted with DMSO (0.5 mL) and filtered through a fritted syringe. The resulting solution was directly purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The pure fractions were combined and lyophilized to afford the formate salt of I-25 (6.1 mg, 0.013 mmol, 10% yield) as a white solid. MS [M+H]⁺=414.2. ¹H NMR (400 MHz, MeCN-d₃) δ 8.79 (s, 1H), 8.23 (s, 1H), 7.54 (d, J=8.4 Hz, 1H), 7.04 (s, 1H), 6.95 (dd, J=8.4, 2.2 Hz, 1H), 4.95 (ddd, J=13.3, 5.2, 2.4 Hz, 1H), 4.91-4.84 (m, 1H), 4.27 (dd, J=16.7, 4.1 Hz, 1H), 4.19 (dd, J=16.7, 4.9 Hz, 1H), 3.56-3.44 (m, 1H), 3.29-3.17 (m, 1H), 2.85-2.57 (m, 4H), 2.31 (qd, J=13.2, 4.8 Hz, 1H), 2.09-1.97 (m, 2H), 1.96-1.88 (m, 1H), 1.74-1.55 (m, 4H), 1.14-0.96 (m, 9H).

Example 9: 3-(5-(((1S,2S)-2-(bis(cyclopropylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-31)

To a stirred solution of the HCl salt of I-12 (26 mg, 0.068 mmol) and NaBH(OAc)₃ (43.5 mg, 0.205 mmol) in DMF (1 mL) was added cyclopropanecarbaldehyde (9-1a, 0.02 mL, 0.2 mmol) and the resulting mixture was stirred overnight at room temperature. The reaction mixture was then concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0% to 10% Et₃N in EtOAc to afford I-31 (12.6 mg, 0.0272 mmol, 40% yield) as a white powder. MS [M+H]⁺=452.3. ¹H NMR (400 MHz, DCM-d₂) δ 8.24 (s, 1H), 7.68 (d, J=8.4 Hz, 1H), 7.17 (s, 1H), 7.10-7.00 (m, 1H), 5.22-5.02 (m, 1H), 4.62 (s, 1H), 4.41-4.21 (m, 2H), 3.70-3.46 (m, 1H), 2.89-2.72 (m, 2H), 2.66-2.23 (m, 5H), 2.20-2.10 (m, 1H), 1.92 (s, 2H), 1.84-1.63 (m, 3H), 1.54 (s, 1H), 0.97-0.82 (m, 2H), 0.45 (s, 4H), 0.23-0.01 (m, 4H).

Example 10: 3-(5-(((1S,2S)-2-(4,4-difluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-8)

3,3-difluoropentane-1,5-diyl dimethanesulfonate 10-1a was prepared as described in U.S. Patent US2017/145026 A1.

To a stirred solution of the HCl salt of I-12 (26 mg, 0.068 mmol) and i-Pr₂NEt (0.05 mL, 0.3 mmol) in DMF (0.7 mL) was added 10-1a (30.4 mg, 0.103 mmol) and the resulting mixture was stirred at 80° C. for 72 h. The reaction mixture was then allowed to cool to room temperature and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 10% Et₃N in EtOAc to afford I-8 (2.4 mg, 5.3 μmol, 8% yield) as a white solid. MS [M+H]⁺=448.1. ¹H NMR (400 MHz, DCM-d₂) δ 8.03 (s, 1H), 7.68-7.57 (m, 1H), 6.99-6.84 (m, 2H), 5.04 (ddd, J=13.4, 5.3, 2.0 Hz, 1H), 4.60-4.50 (m, 1H), 4.31-4.17 (m, 2H), 3.03-2.89 (m, 1H), 2.84-2.68 (m, 2H), 2.63-2.47 (m, 4H), 2.34-2.18 (m, 1H), 2.15-2.07 (m, 1H), 2.03-1.79 (m, 5H), 1.75-1.62 (m, 3H), 1.53-1.41 (m, 2H).

Example 11: 1-((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-methylpiperidine-4-carbonitrile (I-38)

Step 1. 4-hydroxy-2-(2-hydroxyethyl)-2-methylbutanenitrile (11-3a)

To a stirred solution of propiononitrile (11-1a, 1.30 mL, 182 mmol) and Bu₄NI (0.67 g, 1.8 mmol) in THF (30 mL) under an atmosphere of nitrogen was added LDA (12.1 mL, 18.2 mmol, 1.5M in cyclohexane) dropwise at −78° C. and the resulting mixture was stirred for 30 min at −78° C., warmed to room temperature and then stirred for additional 30 min. The reaction mixture was cooled to −78° C. and (2-chloroethoxy)trimethylsilane (11-2a, 2.9 mL, 18 mmol) was added dropwise and the resulting mixture was then warmed to room temperature and stirred for 1 h. The reaction mixture was then cooled to −78° C. and another portion of LDA (12.1 mL, 18.2 mmol, 1.5 M in cyclohexane) was added dropwise. The resulting mixture was stirred for 30 min at −78° C., and then warmed to room temperature and stirred for additional 30 min. The reaction mixture was cooled to −78° C. and a second equivalent of (2-chloroethoxy)trimethylsilane (11-2a, 2.9 mL, 18 mmol) was added dropwise and the reaction mixture was allowed to warm room temperature and then stirred for 3 h. The reaction mixture was quenched with sat. aq. NH₄Cl and extracted with Et₂O (×3). The combined organic phases were concentrated to dryness. The crude product was dissolved in THF (10 mL) and 0.5 M HCl (5 mL) was added. The resulting mixture was stirred for 3 h at room temperature. The reaction mixture was then quenched with sat. aq. NaHCO₃ and extracted with EtOAc (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford 11-3a (250 mg, 1.75 mmol, 10% yield) as a colorless oil. ¹H NMR (400 MHz, Chloroform-d) δ 3.95-3.84 (m, 4H), 2.23 (s, 2H), 2.05-1.95 (m, 2H), 1.93-1.84 (m, 2H), 1.43 (s, 3H).

Step 2. 3-cyano-3-methylpentane-1,5-diyl dimethanesulfonate (11-4a)

To a stirred solution of 11-3a (260 mg, 1.83 mmol) and i-Pr₂NEt (1.3 mL, 7.3 mmol) in DCM (3 mL) was added methanesulfonyl chloride (0.28 mL, 3.6 mmol) at 0° C. and the resulting mixture was stirred for 2 h at 0° C. and then at room temperature for 3 hours. The reaction mixture was then quenched with water (2 mL) and extracted with DCM (×2). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness to afford crude 11-4a as a yellow oil. The crude product was carried onto the next step without further purification. ¹H NMR (400 MHz, DCM-d₂) δ 4.36 (t, J=6.4 Hz, 4H), 3.01 (s, 6H), 2.12 (dt, J=14.7, 6.5 Hz, 2H), 1.99 (dt, J=14.7, 6.4 Hz, 2H), 1.41 (s, 3H).

Step 3. 1-((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-methylpiperidine-4-carbonitrile (I-38)

To a stirred solution of HCl salt of I-12 (500 mg, 1.32 mmol), Bu₄NI (49 mg, 0.13 mmol) and i-Pr₂NEt (1.1 mL, 6.6 mmol) in DMF (3 mL) was added 11-4a (433 mg, 1.45 mmol) and the reaction mixture was stirred for 5 days at 80° C. The reaction mixture was concentrated to dryness. Crude product was purified by silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1) in DCM to afford I-38 (104.7 mg, 0.232 mmol, 18% yield) as a light beige solid. MS [M+H]⁺=451.1. ¹H NMR (400 MHz, DCM-d₂) 8.23 (s, 1H), 7.71-7.66 (m, 1H), 7.09-6.90 (m, 2H), 5.10 (ddd, J=13.3, 5.2, 3.1 Hz, 1H), 4.62 (s, 1H), 4.41-4.19 (m, 2H), 3.10-2.75 (m, 4H), 2.48-2.22 (m, 3H), 2.21-2.11 (m, 1H), 2.11-1.96 (m, 2H), 1.95-1.81 (m, 2H), 1.80-1.64 (m, 3H), 1.63-1.40 (m, 4H), 1.34 (s, 3H).

Example 12: 3-(5-(((1S,2S)-2-(benzylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-20)

To a stirred solution of the HCl salt of I-12 (50 mg, 0.13 mmol) and benzaldehyde (0.02 mL, 0.2 mmol) in anhydrous 2,2,2-trifluoroethanol (1.2 mL) under an atmosphere of nitrogen was added Ti(i-OPr)₄ (0.04 mL, 0.1 mmol) and the resulting mixture was stirred at room temperature overnight. NaBH₄ (7.5 mg, 0.20 mmol) was then added in one portion and the resulting mixture was stirred for an additional 3 h at room temperature. The reaction mixture was quenched with aq. 1 M formic acid (0.2 mL) and filtered. The resulting solution was directly purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The pure fractions were combined and lyophilized to afford I-20 (30.2 mg, 0.062 mmol, 47% yield) as a white powder. MS [M+H]⁺=434.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.97 (s, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.38-7.26 (m, 4H), 7.23-7.16 (m, 1H), 7.13 (dd, J=6.6, 2.1 Hz, 1H), 7.00 (dt, J=8.4, 2.6 Hz, 1H), 5.07 (ddd, J=13.2, 5.1, 3.2 Hz, 1H), 4.71-4.55 (m, 1H), 4.43-4.16 (m, 2H), 3.80-3.67 (m, 2H), 3.14-3.04 (m, 1H), 2.91 (ddd, J=17.4, 13.6, 5.4 Hz, 1H), 2.64-2.54 (m, 1H), 2.45-2.31 (m, 1H), 2.20-2.08 (m, 1H), 2.03-1.94 (m, 1H), 1.93-1.83 (m, 1H), 1.81-1.59 (m, 3H), 1.57-1.45 (m, 1H).

Example 13: 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (I-15)

Step 1. tert-butyl ((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)carbamate (13-2a)

To 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-1d, 1600 mg, 4.95 mmol), tert-butyl ((1S,2S)-2-hydroxycyclohexyl)carbamate (13-1a, 1066 mg, 4.95 mmol), NiCl₂(glyme) (54 mg, 0.25 mmol), dtbbpy (66 mg, 0.25 mmol), Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (56 mg, 0.050 mmol), quinuclidine (55 mg, 0.50 mmol) and K₂CO₃ (684 mg, 4.95 mmol) under an atmosphere of nitrogen was added MeCN (30 mL) and the resulting mixture was then stirred vigorously for 70 h under irradiation of blue LED light at room temperature. The reaction mixture was then diluted with DCM (20 mL), filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 3% MeOH in DCM to afford 13-2a (282 mg, 0.616 mmol, 12% yield) as a white powder. MS [M-tBu+H]⁺=402.3. ¹H NMR (400 MHz, Chloroform-d) δ 7.97 (s, 1H), 7.82-7.70 (m, 1H), 7.13-6.92 (m, 2H), 5.19 (dd, J=13.2, 5.2 Hz, 1H), 4.56 (t, J=7.4 Hz, 1H), 4.42 (d, J=15.8 Hz, 1H), 4.27 (d, J=15.8 Hz, 1H), 4.23-4.16 (m, 1H), 3.78-3.66 (m, 1H), 2.95-2.74 (m, 2H), 2.33 (qd, J=13.0, 5.0 Hz, 1H), 2.25-2.16 (m, 2H), 2.14-2.04 (m, 2H), 1.85-1.74 (m, 1H), 1.46-1.33 (m, 3H), 1.41 (s, 9H).

Step 2. 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (I-15)

To a stirred solution of tert-butyl ((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)carbamate (13-2a, 282 mg, 0.616 mmol) in THF (2 mL) was added 4 M HCl in dioxane (1.0 mL, 4.0 mmol) and the resulting mixture was stirred for 2 h at 60° C. Formation of white precipitate was observed. The reaction mixture was diluted with Et₂O (10 mL) and filtered. The precipitate was washed with Et₂O (×4) and then dried under reduced pressure to afford the hydrochloride salt of I-15 (242 mg, 0.614 mmol, 100% yield) as a white solid. MS [M+H]⁺=358.1. ¹H NMR (400 MHz, D₂O) δ 7.75 (d, J=8.5 Hz, 1H), 7.24 (s, 1H), 7.17 (dd, J=8.5, 2.1 Hz, 1H), 5.12 (dd, J=13.3, 5.2 Hz, 1H), 4.56 (dd, J=17.7, 1.8 Hz, 1H), 4.51-4.38 (m, 2H), 3.45 (ddd, J=12.1, 9.8, 4.3 Hz, 1H), 2.99-2.82 (m, 2H), 2.51 (qd, J=13.0, 5.4 Hz, 1H), 2.39-2.14 (m, 3H), 1.84 (t, J=10.1 Hz, 2H), 1.60 (qd, J=12.5, 4.0 Hz, 1H), 1.49-1.29 (m, 3H).

Alternatively, Conversion of 1-1d to I-15 was Also Achieved Via the Following Synthetic Procedure:

Step 1. 3-(5-bromo-1-oxoisoindolin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (31-3a)

To a stirred solution of 1-1d (10.0 g, 30.9 mmol) and DBU (6.9 mL, 46 mmol) in DMF (95 mL) was added SEMCl (6.6 mL, 37 mmol) at 0° C. and the resulting mixture was allowed to warm to room temperature and then stirred for 5 h. An additional portion of DBU (3.5 mL, 23 mmol) and SEMCl (3.3 mL, 19 mmol) was added and stirring was continued for additional 2 h. The reaction mixture was then quenched with sat. aq. NH₄Cl (250 mL) and extracted with EtOAc (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was dissolved in a minimal amount of EtOAc (˜50 mL) and Et₂O:heptane (v/v=1:2, 400 mL) was added. The resulting cloudy solution was left standing at −5° C. overnight. The formed precipitate was filtered, washed with heptane (×3), and dried under vacuum to afford 31-3a (11.53 g, 25.4 mmol, 82% yield) as an off-white solid. MS [M+H]⁺=453.4. ¹H NMR (400 MHz, Chloroform-d) δ 7.75 (d, J=8.6 Hz, 1H), 7.66-7.61 (m, 2H), 5.37-5.09 (m, 3H), 4.48 (d, J=16.2 Hz, 1H), 4.32 (d, J=16.2 Hz, 1H), 3.74-3.50 (m, 2H), 3.11-2.98 (m, 1H), 2.94-2.83 (m, 1H), 2.33 (qd, J=13.2, 4.7 Hz, 1H), 2.24-2.15 (m, 1H), 0.97-0.90 (m, 2H), 0.00 (s, 9H).

Step 2. tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)carbamate (13-3a)

To a stirred solution of 31-3a (5.00 g, 11.0 mmol), 13-1a (2.37 g, 11.0 mmol), NiCl₂(glyme) (0.121 g, 0.551 mmol), dtbbpy (0.148 g, 0.551 mmol) and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (0.124 g, 0.110 mmol) in degassed MeCN (24 mL) under an atmosphere of nitrogen was added 2,2,6,6-tetramethylpiperidine (40-2, 1.9 mL, 11 mmol) and the resulting mixture was stirred vigorously for 18 h under irradiation of blue LED lights at room temperature. The reaction mixture was then filtered and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford 13-3a (5.50 g, 9.36 mmol, 85% yield) as a yellow foam. MS [M−H]⁻=586.7.

Step 3. 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15)

To a solution of 13-3a (4.88 g, 8.30 mmol) in DCM (60 mL) under an atmosphere of nitrogen was added methanesulfonic acid (2.2 mL, 33 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was cooled to 0° C. and triethylamine (9.3 mL, 66 mmol) and N1,N2-dimethylethane-1,2-diamine (1.1 mL, 10 mmol) were added. The reaction mixture was stirred at rt overnight, quenched with a 50% saturated aqueous sodium hydrogen carbonate, and extracted with DCM:isopropanol (v/v=5:1, ×4). The organic phases were combined, passed through a phase separator and concentrated to dryness. The crude material was suspended in minimal amount of MeCN, sonicated, and filtered. The resulting solid was rinsed with diethyl ether (×3) and dried under vacuum to afford I-12 (1.92 g, 5.37, 65% yield) as a pale yellow solid. MS [M+H]⁺=358.6. ¹H NMR (400 MHz, DMSO-d₆) δ 7.60 (d, J=8.4 Hz, 1H), 7.20 (s, 1H), 7.08 (dd, J=8.4, 2.2 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.43-4.19 (m, 2H), 4.05-3.98 (m, 1H), 2.90 (ddd, J=17.3, 13.6, 5.5 Hz, 1H), 2.82-2.71 (m, 1H), 2.59 (ddd, J=17.3, 4.4, 2.3 Hz, 1H), 2.46-2.30 (m, 1H), 2.11-2.03 (m, 1H), 2.02-1.94 (m, 1H), 1.88-1.80 (m, 1H), 1.73-1.60 (m, 2H), 1.44-1.14 (m, 4H).

Example 14: 3-(5-(((1S,2S)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-22) and 3-(5-(((1S,2S)-2-(dibenzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-34)

To a stirred solution of the HCl salt of I-15 (30.2 mg, 0.073 mmol) in DMF (2 mL) and trifluoroethanol (1 mL) was added benzaldehyde (7 μl, 0.07 mmol) and the resulting mixture was stirred overnight at room temperature. NaBH(OAc)₃ (46 mg, 0.22 mmol) was then added and the resulting mixture was stirred overnight. The reaction mixture was quenched by the addition of one drop of formic acid and then concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 10% Et₃N in EtOAc to afford I-22 and I-34 in about 90% purity. Both compounds were repurified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). Fractions containing the desired product were combined and lyophilized to afford I-22 (6.1 mg, 0.012 mmol, 17% yield) as a white solid and I-34 (7.1 mg, 0.012 mmol, 17% yield) as a white solid. MS of I-22 [M+H]+=448.2. ¹H NMR of I-22 (400 MHz, MeCN-d₃) δ 7.61 (d, J=8.4 Hz, 1H), 7.33-7.26 (m, 4H), 7.25-7.19 (m, 1H), 7.12-7.08 (m, 1H), 7.03 (dd, J=8.4, 2.3 Hz, 1H), 5.03 (ddd, J=13.4, 5.2, 1.9 Hz, 1H), 4.42-4.16 (m, 3H), 3.88 (d, J=13.5 Hz, 1H), 3.77 (d, J=13.5 Hz, 1H), 2.88-2.64 (m, 3H), 2.42-2.32 (m, 2H), 2.18-2.02 (m, 3H), 1.79-1.64 (m, 2H), 1.46-1.21 (m, 4H). MS of I-34 [M+H]⁺=538.4. ¹H NMR of I-34 (400 MHz, MeCN-d₃) δ 8.82 (s, 1H), 7.66 (d, J=8.2 Hz, 1H), 7.33-7.06 (m, 12H), 5.05 (dt, J=13.4, 5.0 Hz, 1H), 4.54 (tt, J=9.6, 4.4 Hz, 1H), 4.41-4.16 (m, 2H), 3.75 (d, J=13.7 Hz, 2H), 3.62 (dd, J=14.1, 4.4 Hz, 2H), 2.90-2.62 (m, 3H), 2.53-2.29 (m, 1H), 2.15-2.06 (m, 2H), 2.06-1.97 (m, 1H), 1.78-1.70 (m, 1H), 1.70-1.61 (m, 1H), 1.46 (qt, J=12.8, 3.3 Hz, 1H), 1.39-1.25 (m, 1H), 1.24-1.06 (m, 2H).

Alternative procedure for synthesis of 3-(5-(((1S,2S)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-22)

To a stirred solution of the HCl salt of I-15 (59 mg, 0.150 mmol) and benzaldehyde (0.08 mL, 0.8 mmol) in TFE (1 mL) was added NaBH(OAc)₃ (159 mg, 0.749 mmol) in one portion and the resulting mixture was stirred vigorously for 3 days at room temperature. The reaction mixture was concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in DCM to afford I-22 (43 mg, 0.096 mmol, 64% yield) as a white solid.

Example 15: 3-(5-(((1S,2S)-2-(methylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-36)

Step 1. 3-(5-(((1S,2S)-2-(benzyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-39)

To a stirred solution of I-22 (43 mg, 0.096 mmol) and NaBH(OAc)₃ (30.5 mg, 0.144 mmol) in TFE (1 mL) was added 37% formaldehyde in H₂O (0.014 mL, 0.19 mmol) and the resulting mixture was stirred at room temperature overnight. The reaction mixture was then diluted with DCM (5 mL), washed with sat. aq. NaHCO₃, dried over Na₂SO₄, filtered, and concentrated to dryness to afford I-39 (44 mg, 0.094 mmol, 98% yield) as a white powder. The product was carried onto the next step without further purification. MS [M+H]⁺=462.1.

Step 2. 3-(5-(((1S,2S)-2-(methylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-36)

To a stirred solution of I-39 (44 mg, 0.095 mmol) in DMF (1 mL) under an atmosphere of nitrogen was added Pd/C (10% wt, 10 mg, 9.5 μmol), followed by triethylsilane (0.05 mL, 0.3 mmol) and the resulting mixture was stirred at room temperature overnight. The reaction mixture was filtered through a pad of Celite® and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in DCM to afford I-36 (14.7 mg, 0.039 mmol, 41% yield) as a white powder. MS [M+H]⁺=372.1. ¹H NMR (400 MHz, DCM-d₂) δ 7.67 (dd, J=8.3, 1.6 Hz, 1H), 7.09-6.94 (m, 2H), 5.07 (dt, J=13.3, 5.1 Hz, 1H), 4.35-4.06 (m, 3H), 2.87-2.68 (m, 2H), 2.62 (dddd, J=10.8, 8.7, 4.2, 2.0 Hz, 1H), 2.38 (d, J=1.7 Hz, 3H), 2.32-2.18 (m, 1H), 2.16-2.01 (m, 3H), 1.80-1.65 (m, 2H), 1.42-1.10 (m, 5H).

Example 16: 3-(5-(((1S,2S)-2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-14)

To a stirred solution of the HCl salt of I-15 (80 mg, 0.18 mmol) and NaBH(OAc)₃ (115 mg, 0.542 mmol) in DMF (2 mL) was added acetaldehyde (2-1, 0.03 mL, 0.5 mmol) and the resulting mixture was stirred for 2 hours at room temperature. The reaction mixture was then concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 10% Et₃N in EtOAc to afford I-14 (30.1 mg, 0.073 mmol, 29% yield) as a white powder. MS [M+H]⁺=414.0. ¹H NMR (400 MHz, DCM-d₂) δ 8.78-8.37 (m, 1H), 7.73 (d, J=8.3 Hz, 1H), 7.16-6.93 (m, 2H), 5.18 (dd, J=13.3, 5.2 Hz, 1H), 4.46-4.24 (m, 3H), 2.94-2.77 (m, 3H), 2.70-2.51 (m, 4H), 2.43-2.29 (m, 1H), 2.28-2.16 (m, 2H), 1.94-1.71 (m, 3H), 1.48-1.23 (m, 4H), 0.97 (t, J=7.1 Hz, 6H).

Example 17: 3-(5-(((1S,2S)-2-(isobutylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-7)

To a stirred solution of the HCl salt of I-15 (50 mg, 0.13 mmol) and NaBH(OAc)₃ (40.4 mg, 0.190 mmol) in DMF (1.5 mL) under an atmosphere of nitrogen was added isobutyraldehyde (17-1a, 0.014 mL, 0.15 mmol) in one portion and the resulting mixture was stirred for 40 min at room temperature. The reaction mixture was then concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 5 mM NH₄OH) and collected into tubes containing formic acid (2 drops). The fractions containing desired product were combined and lyophilized to afford the formate salt of I-7 (17.3 mg, 0.037 mmol, 29% yield) as a white solid. MS [M+H]⁺=414.2. ¹H NMR (400 MHz, DCM-d₂) δ 8.32 (s, 1H), 7.97-7.39 (br s, 1H), 7.72-7.62 (m, 1H), 7.12-6.97 (m, 2H), 5.02 (dt, J=13.2, 5.6 Hz, 1H), 4.56 (dtd, J=13.9, 9.8, 4.1 Hz, 1H), 4.34-4.21 (m, 2H), 3.05-2.93 (m, 1H), 2.81-2.62 (m, 4H), 2.32-2.03 (m, 4H), 1.99-1.86 (m, 1H), 1.84-1.71 (m, 2H), 1.66-1.52 (m, 1H), 1.43-1.21 (m, 3H), 0.96-0.89 (m, 6H).

Example 18: 3-(5-(((3S,4S)-3-(benzylamino)tetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-21)

Step 1. tert-butyl ((3S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)tetrahydro-2H-pyran-3-yl)carbamate (18-2a)

To 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-1d, 550 mg, 1.70 mmol), tert-butyl ((3S,4S)-4-hydroxytetrahydro-2H-pyran-3-yl)carbamate (18-1a, 499 mg, 2.30 mmol), NiCl₂(glyme) (19 mg, 0.085 mmol), dtbbpy (23 mg, 0.085 mmol), Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (19 mg, 0.017 mmol), quinuclidine (18.9 mg, 0.170 mmol) and K₂CO₃ (235 mg, 1.70 mmol) under an atmosphere of nitrogen was added MeCN (15 mL) and the resulting mixture was degassed by bubbling nitrogen during sonication for 40 min. The reaction mixture was then stirred overnight under irradiation of blue LED light at room temperature for 120 h. The reaction mixture was diluted with DCM, filtered through a short pad of Celite®, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 3% MeOH in DCM to afford 18-2a (143 mg, 0.305 mmol, 18% yield) as a white solid. MS [M-tBu+H]⁺=404.2.

Step 2. 3-(5-(((3S,4S)-3-aminotetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione Hydrochloride (I-40)

To a stirred solution of 18-2a (143 mg, 0.311 mmol) in THF (2 mL) was added 4 M HCl in dioxane (1.0 mL, 4.0 mmol) and the resulting mixture was stirred for 3 h at 60° C. Formation of a white precipitate was observed. The reaction mixture was diluted with Et₂O and filtered. The obtained precipitate was washed with Et₂O (4×) and dried under reduced pressure to afford the hydrochloride salt of I-40 (127 mg, 0.305 mmol, 96% yield) as a white solid. The product was carried onto the next step without further purification MS [M+H]⁺=360.2.

Step 3. 3-(5-(((3S,4S)-3-(benzylamino)tetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-21)

To a stirred solution of I-40 (40 mg, 0.10 mmol) and benzaldehyde (0.015 mL, 0.15 mmol) in anhydrous TFE (1.2 mL) under atmosphere of nitrogen was added Ti(i-OPr)₄ (0.03 mL, 0.1 mmol) and the resulting mixture was stirred at room temperature overnight. NaBH₄ (6 mg, 0.2 mmol) was then added in one portion and the resulting mixture was stirred for 2 h at room temperature. The reaction mixture was quenched with H₂O (0.1 mL), stirred for 10 minutes, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing product were combined and lyophilized to afford the formate salt of I-21 (4.4 mg, 0.0087 mmol, 9% yield) as a white solid. MS [M+H]⁺=450.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.97 (s, 1H), 8.26 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.34-7.28 (m, 4H), 7.25-7.18 (m, 2H), 7.09 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.0 Hz, 1H), 4.53-4.45 (m, 1H), 4.36 (dd, J=17.2, 12.2 Hz, 1H), 4.25 (dd, J=17.2, 8.5 Hz, 1H), 3.88 (dd, J=11.8, 4.5 Hz, 1H), 3.84-3.76 (m, 3H), 3.56-3.44 (m, 1H), 3.22 (dd, J=11.4, 8.1 Hz, 1H), 2.91 (ddd, J=17.2, 13.6, 5.4 Hz, 1H), 2.67 (td, J=7.8, 4.1 Hz, 1H), 2.63-2.55 (m, 1H), 2.45-2.32 (m, 1H), 2.19-2.09 (m, 1H), 2.03-1.94 (m, 1H), 1.52 (dtd, J=13.2, 9.2, 4.1 Hz, 1H).

Example 19: 3-(1-oxo-5-(((1S,2S)-2-(((1-(trifluoromethyl)cyclopropyl)methylamino)cyclohexyl)oxy) isoindolin-2-yl)piperidine-2,6-dione (I-11)

Step 1. tert-butyl ((1S,2S)-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclohexyl)carbamate (19-2a)

To 5-bromoisobenzofuran-1(3H)-one (19-1a, 500 mg, 2.35 mmol), tert-butyl ((1S,2S)-2-hydroxycyclohexyl)carbamate (13-1a, 505 mg, 2.35 mmol), NiCl₂(glyme) (26 mg, 0.12 mmol), dtbbpy (32 mg, 0.12 mmol), Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (26 mg, 0.023 mmol), quinuclidine (26 mg, 0.24 mmol) and K₂CO₃ (324 mg, 2.35 mmol) was added MeCN (30 mL) and the resulting mixture was stirred vigorously for 22 h under irradiation of blue LED light at room temperature. The reaction mixture was then concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford 19-2a (732 mg, 1.93 mmol, 82% yield) as a white powder. MS [M-tBu+H]⁺=292.1.

Step 2. 5-(((1S,2S)-2-aminocyclohexyl)oxy)isobenzofuran-1(3H)-one CF₃CO₂H Salt (19-3a)

To a stirred solution 19-2a (700 mg, 1.85 mmol) in DCM (0.6 mL) was added CF₃CO₂H (0.6 mL, 7.8 mmol) and the resulting mixture was stirred at room temperature overnight. The reaction mixture was then concentrated to dryness by azeotroping with heptane (3×) to afford the trifluoroacetate salt of 19-3a (710 mg, 1.83 mmol, 99% yield) as viscous yellow oil. The crude product was carried onto the next step without further purification. MS [M+H]⁺=247.9.

Step 3. 5-(((1S,2S)-2-(((1-(trifluoromethyl)cyclopropyl)methyl)amino)cyclohexyl)oxy)isobenzofuran-1(3H)-one (19-5a)

To a stirred solution of 19-3a (300 mg, 0.830 mmol) and i-Pr₂NEt (0.43 mL, 2.5 mmol) in DMF (2.0 mL) was added 1-(bromomethyl)-1-(trifluoromethyl)cyclopropane (19-4a, 0.2 mL, 1 mmol) and the resulting mixture was stirred at 85° C. for 5 h. The reaction mixture was then concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0-100% EtOAc (with 1% NEt₃) in heptane to afford 19-5a (53 mg, 0.14 mmol, 17% yield) as a white powder. MS [M+H]⁺=370.2. ¹H NMR (400 MHz, DCM-d₂) δ 7.73 (d, J=8.5 Hz, 1H), 7.02 (dd, J=8.5, 2.2 Hz, 1H), 6.95 (s, 1H), 5.20 (s, 2H), 4.10 (ddd, J=10.0, 8.4, 4.2 Hz, 1H), 2.84 (q, J=13.1 Hz, 2H), 2.71 (ddd, J=10.6, 8.6, 4.1 Hz, 1H), 2.19-2.06 (m, 1H), 2.04-1.95 (m, 1H), 1.82-1.59 (m, 3H), 1.42-1.23 (m, 3H), 1.21-1.09 (m, 1H), 0.92-0.85 (m, 2H), 0.69-0.61 (m, 2H).

Step 4. Ethyl 2-(chloromethyl)-4-(((1S,2S)-2-(((1-(trifluoromethyl)cyclopropyl)methyl)amino)cyclohexyl)oxy)benzoate (19-6a)

To a stirred solution of 19-5a (53 mg, 0.14 mmol) in EtOH (0.5 mL) was added SOCl₂ (0.06 mL, 0.9 mmol) and the resulting mixture was stirred overnight at 70° C. An additional portion of SOCl₂ (0.02 mL, 0.3 mmol) was added and the reaction mixture was stirred for another 7 h at 70° C. The reaction mixture was then allowed to cool to room temperature and concentrated to dryness by azeotroping with heptane (2×) to afford crude 19-6a as a brown oil which was used in the next step without further purification. MS [M+H]⁺=435.7.

Step 5. 3-(1-oxo-5-(((1S,2S)-2-(((1-(trifluoromethyl)cyclopropyl)methyl)amino)cyclohexyl)oxy) isoindolin-2-yl)piperidine-2,6-dione (I-11)

To a stirred solution of crude (19-6a) (˜0.14 mmol) and i-Pr₂NEt (0.10 mL, 0.57 mmol) in DMF (0.5 mL) was added 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 47 mg, 0.29 mmol) and the resulting mixture was stirred at 85° C. for 48 h. The reaction mixture was cooled to room temperature and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 10% Et₃N in EtOAc to afford I-11 (8.3 mg, 0.017 mmol, 12% yield over 2 steps) as a white solid. MS [M+H]⁺=480.0. ¹H NMR (400 MHz, DCM-d₂) δ 8.29 (s, 1H), 7.79-7.55 (m, 1H), 7.04-6.95 (m, 2H), 5.11 (ddd, J=13.3, 5.2, 2.9 Hz, 1H), 4.40-4.23 (m, 2H), 4.09 (td, J=9.2, 4.3 Hz, 1H), 2.91-2.77 (m, 4H), 2.77-2.65 (m, 1H), 2.32 (qd, J=12.8, 5.8 Hz, 1H), 2.22-2.07 (m, 2H), 2.05-1.94 (m, 1H), 1.81-1.63 (m, 3H), 1.44-1.13 (m, 4H), 0.94-0.86 (m, 2H), 0.70-0.64 (m, 2H).

Example 20: 3-(1-oxo-5-(((1S,2S)-2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-23)

Step 1. 5-(((1S,2S)-2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)isobenzofuran-1(3H)-one (20-2a)

(S)-1-phenylethyl methanesulfonate 20-1a was prepared from (S)-1-phenylethanol as reported in U.S. Patent US2018/9796 A1, 2018 (also see Example 19, Steps 1 and 2).

To a stirred solution of 19-3a (170 mg, 0.471 mmol) and i-Pr₂NEt (0.41 mL, 2.35 mmol) in MeCN (1 mL) was added a solution of (S)-1-phenylethyl methanesulfonate 20-1a (451 mg, 2.25 mmol) in MeCN (1 mL) and the resulting mixture was stirred for 72 h at room temperature. The reaction mixture was then concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc (with 1% Et₃N) in heptane to afford 20-2a (60 mg, 0.17 mmol, 36% yield) as a white powder. MS [M+H]⁺=352.0. ¹H NMR (400 MHz, DCM-d₂) δ 7.72 (d, J=8.5 Hz, 1H), 7.38-7.22 (m, 5H), 6.98 (dd, J=8.5, 2.2 Hz, 1H), 6.83 (s, 1H), 5.17 (d, J=2.1 Hz, 2H), 4.13 (ddd, J=9.3, 7.8, 4.0 Hz, 1H), 3.91 (q, J=6.6 Hz, 1H), 2.54 (ddd, J=9.2, 7.8, 4.1 Hz, 1H), 2.11-2.00 (m, 2H), 1.86 (s, 1H), 1.75-1.59 (m, 2H), 1.41-1.12 (m, 4H) 1.27 (d, J=6.7 Hz, 3H).

Step 2. Ethyl 2-(chloromethyl)-4-(((1S,2S)-2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)benzoate (20-3a)

To a stirred solution of 20-2a (60 mg, 0.17 mmol) in EtOH (1.0 mL) was added SOCl₂ (0.074 mL, 1.0 mmol) and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was then cooled to room temperature and concentrated to dryness by azeotroping with PhMe (×3) to afford crude 20-3a as a pale brown solid, which was carried onto the next step without further purification. MS [M+H]⁺=416.1.

Step 3. 3-(1-oxo-5-(((1S,2S)-2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-23)

To a stirred solution of crude 20-3a (˜0.17 mmol) and i-Pr₂NEt (0.149 mL, 0.855 mmol) in DMF (1 mL) was added 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 37.2 mg, 0.291 mmol) and the resulting mixture was stirred at 85° C. overnight and then at 110° C. for 4 h. The reaction mixture was allowed to cool to room temperature and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 10% Et₃N in EtOAc to afford I-23 (33.3 mg, 0.071 mmol, 42% yield over 2 steps) as an off-white powder. MS [M+H]⁺=462.2. ¹H NMR (400 MHz, DCM-d₂) δ 7.67 (d, J=8.4 Hz, 1H), 7.37-7.19 (m, 5H), 6.98-6.90 (m, 2H), 5.09 (ddd, J=13.4, 5.2, 2.6 Hz, 1H), 4.34-4.20 (m, 2H), 4.15-4.04 (m, 1H), 3.94-3.84 (m, 1H), 2.88-2.72 (m, 2H), 2.54-2.45 (m, 1H), 2.37-2.22 (m, 1H), 2.20-2.09 (m, 1H), 2.08-1.96 (m, 2H), 1.70-1.55 (m, 2H), 1.36-1.09 (m, 4H), 1.25 (d, J=6.7 Hz, 3H).

Example 21: 3-(5-(((1S,2S)-2-hydroxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-27)

Step 1. Methyl 2-(bromomethyl)-4-iodobenzoate (21-2a)

Into a 3 L three-necked round-bottom flask, purged and maintained under an inert atmosphere of nitrogen, was placed methyl 4-iodo-2-methylbenzoate (21-1a, 170 g, 616 mmol), MeCN (1.0 L), AIBN (10.1 g, 61.5 mmol), and NBS (131.6 g, 739.2 mmol). The resulting solution was stirred overnight at 80° C. and then cooled to room temperature. The solids were filtered off and the resulting mixture was concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 10% EtOAc/petroleum ether. The collected fractions were concentrated under vacuum to afford 21-2a (50.0 g, 141 mmol, 23% yield) as a white solid. ¹H NMR (300 MHz, DMSO-d₆) δ 8.04-8.01 (m, 1H), 7.88-7.81 (m, 1H), 7.67-7.59 (m, 1H), 4.96 (s, 2H), 3.87 (s, 3H).

Step 2. 3-(5-iodo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (21-3a)

1 L three-necked round-bottom flask was charged with 21-2a (50 g, 141 mmol), 3 aminopiperidine-2,6-dione CF₃CO₂H salt (I-1c, 34.18 g, 141.2 mmol), DMF (500 mL), and Et₃N (42.4 g, 419 mmol) and the resulting solution was stirred for 48 h at 60° C. The reaction mixture was cooled to room temperature and quenched by the addition of 500 mL of water/ice. The pH value of the solution was adjusted to pH=5 with 1 M aq. HCl. The solids were collected by filtration, washed with 3×500 mL of EtOAc and dried under vacuum to afford 21-3a (13 g, 35 mmol, 25% yield) as a grey solid. [M+H]⁺=371.0. ¹H NMR (300 MHz, DMSO-d₆) δ 11.00 (s, 1H), 8.06 (s, 1H), 7.90 (d, J=7.8 Hz, 1H), 7.52 (d, J=7.8 Hz, 1H), 5.14-5.08 (m, 1H), 4.47-4.28 (m, 2H), 2.97-2.85 (m, 1H), 2.73-2.01 (m, 2H), 1.98-1.20 (m, 1H).

Step 3. 3-(5-(((1S,2S)-2-hydroxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-27)

To a stirred suspension of 3-(5-iodo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (21-3a, 30 mg, 0.081 mmol), quinuclidine (1 mg, 8 μmol), Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (1 mg, 0.8 μmol) and K₂CO₃ (11.2 mg, 0.09 mmol) in MeCN (1.5 mL) was added (1S,2S)-cyclohexane-1,2-diol (21-4a, 24 mg, 0.20 mmol), followed by an aliquot of Ni catalyst solution (0.025 M, 0.16 mL, 4 μmol), prepared by dissolving NiCl₂(glyme) (22 mg, 0.1 mmol) and dtbbpy (27 mg, 0.1 mmol) in MeCN (4 mL). The resulting mixture was degassed by bubbling nitrogen into the solution under sonication for 30 min and then stirred vigorously for 5 days under the irradiation of blue LED light at room temperature. The reaction mixture was diluted with MeCN (4 mL), filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing product were combined and lyophilized to afford I-27 (3.2 mg, 0.0089 mmol, 11% yield) as a white solid. MS [M+H]⁺=359.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.59 (d, J=8.5 Hz, 1H), 7.31-7.14 (m, 1H), 7.06 (dd, J=8.4, 2.2 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.95 (s, 1H), 4.37 (dd, J=17.1, 4.9 Hz, 1H), 4.24 (dd, J=17.1, 4.4 Hz, 1H), 4.20-4.06 (m, 1H), 3.62-3.50 (m, 1H), 2.90 (ddd, J=17.4, 13.6, 5.4 Hz, 1H), 2.65-2.52 (m, 1H), 2.37 (qd, J=13.2, 4.3 Hz, 1H), 2.10-1.91 (m, 2H), 1.91-1.82 (m, 1H), 1.71-1.57 (m, 2H), 1.42-1.18 (m, 4H).

Example 22: 3-(5-(((1R*, 2S*)-2-hydroxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-37)

To a stirred suspension of 3-(5-iodo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (21-3a, 30 mg, 0.081 mmol), Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (1 mg, 0.8 μmol), quinuclidine (1 mg, 8 μmol), K₂CO₃ (1.2 mg, 0.09 mmol) in MeCN (1.5 mL) was added cis-cyclohexane-1,2-diol (22-1a, 24 mg, 0.20 mmol), followed by an aliquot of Ni catalyst solution (0.025 M, 0.16 mL, 4 μmol). prepared by dissolving NiCl₂(glyme) (22 mg, 0.1 mmol) and dtbbpy (27 mg, 0.1 mmol) in MeCN (4 mL). The resulting mixture was degassed by bubbling nitrogen through the mixture under sonication for 30 min and then stirred vigorously for 4 days under irradiation of blue LED light at room temperature. The reaction mixture was diluted with DCM (4 mL), filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were lyophilized to afford I-37 (2.1 mg, 0.0056 mmol, 7% yield) as a white solid. MS [M+H]⁺=359.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.95 (s, 1H) 7.80 (d, J=8.4 Hz, 1H), 7.05 (d, J=8.4, 2.2 Hz, 1H), 7.00 (d, J=2.1 Hz, 1H), 5.20 (dd, J=13.1, 5.2 Hz, 1H), 4.50 (d, J=7.3 Hz, 1H), 4.44 (d, J=15.8 Hz, 1H), 4.29 (d, J=158 Hz, 1H), 3.96 (d, J=7.5 Hz, 1H), 2.98-2.75 (m, 2H), 2.34 (qd, J=13.1, 5.0 Hz, 1H), 2.27-2.17 (m, 1H), 2.07-1.85 (m, 2H), 1.79-1.68 (m, 2H), 1.68-1.62 (m, 2H), 1.46-1.32 (m, 3H).

Example 23: 3-(5-(((1R,2S)-2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-35)

Step 1. tert-butyl ((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl) carbamate (23-2a)

To 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-1d, 800 mg, 2.48 mmol), NiCl₂(glyme) (27 mg, 0.12 mmol), dtbbpy (33 mg, 0.12 mmol), quinuclidine (28 mg, 0.25 mmol), tert-butyl ((1S,2R)-2-hydroxycyclohexyl)carbamate (23-1a, 533 mg, 2.48 mmol) and K₂CO₃ (342 mg, 2.48 mmol) under an atmosphere of nitrogen was added MeCN (15 mL). The resulting mixture was degassed by bubbling nitrogen through the mixture under sonication for 30 min and then stirred vigorously for 100 hours under irradiation of blue LED light at room temperature. The reaction mixture was diluted with DCM, filtered through a pad of Celite®, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 3% MeOH in DCM to afford 23-2a (121 mg, 0.16 mmol, 6.5% yield) as a white solid with a purity of about 60%. The product was carried onto the next step without further purification. MS [M-tBu+H]⁺=402.3.

Step 2. 3-(5-(((1R,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (I-41)

To a stirred solution of 23-2a (121 mg, 0.161 mmol, ˜60% pure) in THF (2 mL) was added 4 M HCl in dioxane (1.0 mL, 4.0 mmol) and the resulting mixture was stirred for 2 h a 60° C. Formation of white precipitate was observed. The reaction mixture was diluted with Et₂O and filtered. The white precipitate was washed with Et₂O (4×) and dried under vacuum to afford the hydrochloride salt of I-41 (64 mg, 0.16 mmol, 94% yield) as a white solid. The product was carried onto the next step without further purification. MS [M+H]⁺=358.1.

Step 3. 3-(5-(((1R,2S)-2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-35)

To a stirred solution of I-41 (50 mg, 0.13 mmol) and acetaldehyde (2-1, 0.03 mL, 0.5 mmol) in anhydrous TFE (2 mL) under and atmosphere of nitrogen was added NaBH(OAc)₃ (103 mg, 0.487 mmol) in one portion and the resulting mixture was stirred for 5 hours at room temperature. The reaction mixture was concentrated to dryness, redissolved in DMF (2 mL) and additional NaBH(OAc)₃ (103 mg, 0.487 mmol) and acetaldehyde (2-1, 0.03 mL, 0.5 mmol) were added. The resulting reaction mixture was stirred for 2.5 h and then quenched with 1 M aq. HCOOH (0.2 mL). The mixture was directly purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing product were combined and lyophilized to afford the formate salt of I-35 (10.9 mg, 0.023 mmol, 14% yield) as a white powder. MS [M+H]⁺=414.3. ¹H NMR (400 MHz, MeCN-d₃) δ 8.37 (s, 1H), 7.66 (d, J=8.4 Hz, 1H), 7.19-7.17 (m, 1H), 7.12 (dt, J=8.4, 2.2 Hz, 1H), 5.10-5.01 (m, 2H), 4.37 (dd, J=16.8, 2.9 Hz, 1H), 4.29 (dd, J=16.8, 3.8 Hz, 1H), 3.18 (d, J=12.5 Hz, 1H), 3.08-2.90 (m, 4H), 2.88-2.65 (m, 2H), 2.40 (qd, J=13.2, 4.9 Hz, 1H), 2.18-1.99 (m, 3H), 1.92-1.82 (m, 2H), 1.63-1.37 (m, 4H), 1.14 (t, J=7.2 Hz, 6H).

Example 24: 3-(5-(((1R,2R)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-43)

Step 1. tert-butyl ((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)carbamate (24-2a)

To a stirred suspension of 1-Id (263 mg, 0.813 mmol), tert-butyl ((1R,2R)-2-hydroxycyclopentyl)carbamate (24-1a, 149 mg, 0.740 mmol), NiCl₂(glyme) (9 mg, 0.04 mmol), dtbbpy (11 mg, 0.042 mmol), and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (10 mg, 9.1 μmol) in MeCN (2.5 mL) under an atmosphere of nitrogen was added 2,2,6,6-tetramethylpiperidine (40-2, 0.13 mL, 0.77 mmol) and the resulting mixture was stirred vigorously overnight under irradiation of blue LED light at room temperature. The reaction mixture was then diluted with EtOAc, filtered through Celite®, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and lyophilized to afford 24-2a as a white solid (30.8 mg, 0.067 mmol, 9% yield). MS [M+H]⁺=444.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (s, 1H), 7.11-6.99 (m, 2H), 5.07 (ddd, J=13.4, 5.2, 1.9 Hz, 1H), 4.72-4.56 (m, 1H), 4.52-4.16 (m, 2H), 3.88 (s, 1H), 2.90 (ddd, J=18.3, 13.6, 5.4 Hz, 1H), 2.60 (dd, J=3.8, 1.8 Hz, 1H), 2.38 (dd, J=13.2, 4.5 Hz, 1H), 2.14-1.83 (m, 3H), 1.83-1.58 (m, 3H), 1.50 (dq, J=13.5, 7.1, 6.5 Hz, 1H), 1.38 (s, 9H).

Step 2. 3-(5-(((1R,2R)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (I-42)

To a stirred solution of 24-2a (30 mg, 0.068 mmol) in THF (1 mL) was added 4 M HCl in dioxane (0.1 mL, 0.4 mmol) and the resulting mixture was stirred at room temperature for 48 h. The reaction mixture was then filtered and the obtained white precipitate was washed with EtOAc multiple times. The solid was then collected to afford the hydrochloride salt of I-42 as a yellow solid (10 mg, 0.025 mmol, 37% yield). MS [M+H]⁺=344.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.97 (s, 1H), 8.31 (s, 3H), 7.66 (d, J=8.4 Hz, 1H), 7.22 (dd, J=4.6, 2.2 Hz, 1H), 7.08 (dt, J=8.4, 2.3 Hz, 1H), 5.14-5.02 (m, 1H), 4.85 (dt, J=7.3, 4.2 Hz, 1H), 4.47-4.23 (m, 2H), 3.68-3.62 (m, 1H), 2.91 (ddd, J=17.3, 13.6, 5.4 Hz, 1H), 2.66-2.56 (m, 1H), 2.44-2.34 (m, 1H), 2.31-2.22 (m, 1H), 2.17-2.06 (m, 1H), 2.06-1.92 (m, 1H), 1.90-1.62 (m, 4H).

Step 3. 3-(5-(((1R,2R)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-43)

To a stirred solution of I-42 (10 mg, 0.026 mmol) and acetaldehyde (2-1, 4 μL, 0.08 mmol) in DMF (0.5 mL) was added sodium triacetoxyborohydride (17 mg, 0.079 mmol) in one portion and the resulting mixture was stirred at room temperature for 8 hours. The reaction mixture was then concentrated to dryness and the crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and lyophilized to afford the formate salt of I-43 as a white solid (7.8 mg, 0.017 mmol, 65% yield). MS [M+H]⁺=400.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.18 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.22 (t, J=1.9 Hz, 1H), 7.05 (dt, J=8.4, 2.0 Hz, 1H), 5.12-5.00 (m, 1H), 4.65 (s, 1H), 4.47-4.19 (m, 2H), 2.90 (ddd, J=18.3, 13.7, 5.4 Hz, 1H), 2.60 (d, J=2.9 Hz, 1H), 2.38 (dd, J=13.1, 4.5 Hz, 1H), 2.07-1.94 (m, 2H), 1.89 (d, J=6.3 Hz, 1H), 1.76-1.58 (m, 3H), 1.55-1.44 (m, 1H), 0.96 (t, J=7.1 Hz, 6H).

Example 25: 3-(5-(((1S,2R)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-10)

Step 1. tert-butyl ((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)carbamate (25-2a)

To a stirred suspension of 1-Id (280 mg, 0.868 mmol), tert-butyl ((1R,2S)-2-hydroxycyclopentyl)carbamate (25-1a, 167 mg, 0.828 mmol), NiCl₂(glyme) (9.5 mg, 0.043 mmol), dtbbpy (11.5 mg, 0.043 mmol), and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (10 mg, 8.8 μmol) in MeCN (2.5 mL) under an atmosphere of nitrogen was added 2,2,6,6-tetramethylpiperidine (40-2, 0.15 mL, 0.89 mmol) and the resulting mixture was stirred overnight under the irradiation of blue LED light at room temperature. The reaction mixture was then diluted with EtOAc, filtered through Celite®, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing desired product were combined and lyophilized to afford 25-2a as a white solid (78.1 mg, 0.174 mmol, 21% yield). MS [M−H]⁻=442.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.12 (s, 1H), 7.02 (d, J=8.3 Hz, 1H), 6.81 (t, J=7.1 Hz, 1H), 5.06 (dd, J=13.3, 5.0 Hz, 1H), 4.75 (s, 1H), 4.45-4.17 (m, 2H), 3.92 (s, 1H), 2.90 (ddd, J=18.3, 13.6, 5.4 Hz, 1H), 2.60 (s, 1H), 2.38 (dd, J=13.2, 4.5 Hz, 1H), 2.07-1.91 (m, 2H), 1.91-1.80 (m, 1H), 1.80-1.62 (m, 3H), 1.62-1.45 (m, 1H), 1.30 (d, J=2.8 Hz, 9H).

Step 2. 3-(5-(((1S,2R)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (I-28)

To a stirred solution of 25-2a (78.1 mg, 0.176 mmol) in THF (1 mL) was added 4 M HCl in dioxane (0.15 mL, 0.60 mmol) and the resulting mixture was allowed to stir at room temperature for 36 h. The reaction mixture was filtered, washed with EtOAc multiple times, and dried under vacuum to afford hydrochloride salt of I-28 as a yellow solid. MS [M+H]⁺=344.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.97 (s, 1H), 8.10 (s, 3H), 7.68 (d, J=8.4 Hz, 1H), 7.23 (t, J=2.5 Hz, 1H), 7.13 (dt, J=8.5, 1.9 Hz, 1H), 5.08 (dd, J=13.3, 5.0 Hz, 1H), 4.88 (s, 1H), 4.50-4.22 (m, 2H), 3.69 (s, 1H), 2.91 (ddd, J=17.7, 13.4, 5.3 Hz, 1H), 2.65-2.57 (m, 1H), 2.38 (dd, J=13.2, 4.5 Hz, 1H), 2.16-1.92 (m, 3H), 1.88-1.73 (m, 3H), 1.73-1.59 (m, 1H).

Step 3. 3-(5-(((1S,2R)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-10)

To a stirred solution of I-28 (50 mg, 0.13 mmol) and acetaldehyde (2-1, 0.02 mL, 0.4 mmol) in DMF (1 mL) was added sodium triacetoxyborohydride (85 mg, 0.40 mmol) in one portion and the resulting mixture was stirred at room temperature for 24 h. The reaction mixture was then concentrated to dryness and the crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and lyophilized to afford the formate salt of I-10 as a white solid (23.1 mg, 0.049 mmol, 37% yield). MS [M+H]⁺=400.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.16 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.16 (d, J=2.1 Hz, 1H), 7.05 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.0 Hz, 1H), 4.88 (s, 1H), 4.50-4.16 (m, 2H), 3.09 (s, 1H), 2.96-2.80 (m, 1H), 2.75-2.62 (m, 5H), 2.64-2.53 (m, 1H), 2.39 (dd, J=13.2, 8.7 Hz, 1H), 2.03-1.67 (m, 5H), 1.67-1.57 (m, 1H), 0.95 (t, J=7.0 Hz, 6H).

Example 26: 3-(5-(((1S,2S)-2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-24)

Step 1. tert-butyl ((1S,2S)-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate (26-1a)

To 5-bromoisobenzofuran-1(3H)-one (19-1a, 1.12 g, 5.26 mmol), tert-butyl ((1S,2S)-2-hydroxycyclopentyl)carbamate (1-1e, 1.06 g, 5.26 mmol), NiCl₂(glyme) (55 mg, 0.25 mmol), dtbbpy (67 mg, 0.25 mmol), Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (56 mg, 0.050 mmol), quinuclidine (55 mg, 0.50 mmol) and K₂CO₃ (688 mg, 4.98 mmol) under an atmosphere of nitrogen was added MeCN (25 mL) and the resulting mixture was stirred vigorously for 30 hours under irradiation of blue LED light at room temperature. The reaction mixture was then diluted with DCM (20 mL), filtered through a pad of Celite R, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford 26-1a (1.38 g, 3.89 mmol, 74% yield) as a white solid. MS [M-tBu+H]⁺=278.1. ¹H NMR (400 MHz, DCM-d₂) δ 7.75 (d, J=8.7 Hz, 1H), 7.24-7.03 (m, 2H), 5.23 (s, 2H), 4.69 (s, 1H), 4.60 (s, 1H), 4.09-3.99 (m, 1H), 2.22-2.02 (m, 2H), 1.91-1.75 (m, 3H), 1.59-1.48 (m, 1H), 1.43 (s, 9H).

Step 2. 5-(((1S,2S)-2-aminocyclopentyl)oxy)isobenzofuran-1 (3H)-one CF₃CO₂H Salt (26-2a)

To a stirred solution of 26-1a (1.9 g, 5.70 mmol) in DCM (5 mL) was added TFA (2.18 mL, 28.5 mmol) and the resulting mixture was stirred at room temperature overnight. The reaction mixture was concentrated to dryness by azeotroping with PhMe (×2) to afford the trifluoroacetate salt of 26-2a (1.97 g, 5.45 mmol, 96% yield) as a white solid. The product was carried onto the next step without further purification. MS [M+H]⁺=234.3.

Step 3. 5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)isobenzofuran-1(3H)-one (26-3a)

To a stirred solution of 26-2a (155.6 mg, 0.448 mmol) and acetaldehyde (2-1, 0.03 mL, 0.5 mmol) in 2,2,2-trifluoroethanol (2 mL) under an atmosphere of nitrogen was added NaBH(OAc)₃ (199 mg, 0.939 mmol) in one portion and the resulting mixture was stirred at room temperature until complete consumption of the starting material was observed. The reaction mixture was diluted with water (20 mL) and basified with sat. aq. NaHCO₃. The aqueous solution was extracted with EtOAc (3×25 mL) and the combined organic phases were washed with brine (20 mL), dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 10% MeOH in DCM (with 0.1% Et₃N modifier) to afford 26-3a (79.5 mg, 0.283 mmol, 63% yield) as a light yellow gum. MS [M+H]⁺=262.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.79 (d, J=8.5 Hz, 1H), 7.07 (dd, J=8.5, 2.2 Hz, 1H), 7.01-6.99 (m, 1H), 5.26 (s, 2H), 4.57 (dt, J=6.4, 3.4 Hz, 1H), 3.31 (td, J=6.7, 3.7 Hz, 1H), 2.82-2.61 (m, 2H), 2.25-2.06 (m, 2H), 1.86-1.73 (m, 3H), 1.54-1.45 (m, 2H), 1.13 (t, J=7.1 Hz, 3H).

Step 4. 5-(((1S,2S)-2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)isobenzofuran-1 (3H)-one (26-4a)

To a stirred solution of 26-3a (79.5 mg, 0.304 mmol) and 1-fluoro-2-iodoethane (140 mg, 0.805 mmol) in DMF (1 mL) was added i-Pr₂NEt (0.15 mL, 0.86 mmol) and the resulting mixture was stirred at 85° C. overnight. The reaction mixture was diluted with EtOAc (40 mL) and washed with 0.5 M aq. LiCl solution (10 mL), sat. aq. NaHCO₃ (10 mL), and brine (10 mL). The combined organic layers were dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 5% MeOH in DCM (with 0.1% Et₃N modifier) to afford 26-4a as a colorless gum (39.8 mg, 0.128 mmol, 42% yield). MS [M+H]⁺=308.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.79 (d, J=8.4 Hz, 1H), 7.22-7.05 (m, 2H), 5.26 (s, 2H), 4.75-4.30 (m, 2H), 3.41 (s, 1H), 2.93-2.74 (m, 2H), 2.73-2.56 (m, 2H), 2.02 (s, 2H), 1.85 (dd, J=25.5, 11.3 Hz, 3H), 1.55 (d, J=11.2 Hz, 2H), 1.08 (s, 3H).

Step 5. Ethyl 2-(chloromethyl)-4-(((1S,2S)-2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)benzoate (26-5a)

To a stirred solution of 26-4a (39.8 mg, 0.129 mmol) in EtOH (1 mL) was added SOCl₂ (0.05 mL, 0.7 mmol) and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was diluted with water (10 mL), neutralized with sat. aq. NaHCO₃, and extracted with EtOAc (3×30 mL). The combined organic phases were then washed with 5% aq. NaHCO₃ solution (10 mL), brine (10 mL), dried over MgSO₄, filtered, and concentrated to dryness to afford the crude 26-5a as a brown-red gum (45.9 mg, 0.114 mmol, 88% yield). MS [M+H]⁺=372.2. ¹H NMR (400 MHz, DCM-d₂) δ 8.00 (d, J=8.8 Hz, 1H), 7.19 (d, J=2.6 Hz, 1H), 6.99 (dd, J=8.8, 2.6 Hz, 1H), 5.07 (s, 2H), 4.64 (d, J=4.9 Hz, 1H), 4.54 (t, J=5.5 Hz, 1H), 4.42 (t, J=5.5 Hz, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.40 (td, J=8.4, 4.1 Hz, 1H), 2.84 (dt, J=24.5, 5.3 Hz, 2H), 2.68 (q, J=7.0 Hz, 2H), 2.10-1.95 (m, 2H), 1.88-1.73 (m, 2H), 1.57 (d, J=16.7 Hz, 2H), 1.41 (t, J=7.1 Hz, 3H), 1.08 (t, J=7.1 Hz, 3H).

Step 6. 3-(5-(((1S,2S)-2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-24)

To a stirred solution of 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 26 mg, 0.16 mmol) and i-Pr₂NEt (0.07 mL, 0.38 mmol) in DMF (0.5 mL) was added a solution of 26-5a (45.9 mg, 0.123 mmol) in DMF (0.5 mL) and the resulting mixture was stirred at 85° C. until complete consumption of starting material was observed by LC-MS analysis. The reaction mixture was diluted with EtOAc (40 mL), washed with sat. aq. NaHCO₃ (3×10 mL), brine (10 mL), dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and lyophilized to afford I-24 as a white solid (6.5 mg, 0.015 mmol, 12% yield). MS [M+H]⁺=418.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.24 (d, J=2.1 Hz, 1H), 7.06 (dt, J=8.5, 2.0 Hz, 1H), 5.07 (dd, J=13.2, 5.1 Hz, 1H), 4.65 (s, 1H), 4.48 (t, J=5.3 Hz, 1H), 4.44-4.19 (m, 3H), 2.98-2.71 (m, 3H), 2.64-2.56 (m, 3H), 2.38 (dd, J=13.2, 4.5 Hz, 1H), 2.11-1.85 (m, 4H), 1.79-1.56 (m, 3H), 1.54-1.42 (m, 1H), 0.98 (t, J=7.1 Hz, 3H).

Example 27: 3-(5-(((1S,2S)-2-((2,2-difluoroethyl)(ethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-1)

Step 1. 5-(((1S,2S)-2-((2,2-difluoroethyl)(ethyl)amino)cyclopentyl)oxy)isobenzofuran-1(3H)-one (27-2a)

To a stirred solution of 27-1a (47.1 mg, 0.180 mmol) and 1,1-difluoro-2-iodoethane (32 mg, 0.17 mmol) in DMF (1 mL) was added i-Pr₂NEt (0.07 mL, 0.4 mmol) and the resulting mixture was stirred at room temperature overnight. The reaction mixture was then heated to 85° C. and stirred until complete consumption of starting material was observed by LC-MS analysis. The solution was diluted with EtOAc (40 mL) and washed with 0.5 M aq. LiCl solution (10 mL), sat. aq. NaHCO₃ solution (10 mL), and brine (10 mL). The combined organic layers were dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 5% MeOH in DCM (0.1% Et₃N modifier) to afford the 27-2a as a colorless gum (28.2 mg, 0.077 mmol, 43% yield). MS [M+H]⁺=326.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.79 (d, J=8.2 Hz, 1H), 7.13-7.06 (m, 2H), 5.79 (tt, J=56.4, 4.3 Hz, 1H), 5.26 (s, 2H), 4.72-4.56 (m, 1H), 3.42 (dt, J=13.5, 9.0 Hz, 1H), 2.97-2.82 (m, 3H), 2.78-2.64 (m, 1H), 2.23-1.97 (m, 2H), 1.92-1.74 (m, 3H), 1.55 (d, J=11.9 Hz, 2H), 1.09 (t, J=7.1 Hz, 2H).

Step 2. Ethyl 2-(chloromethyl)-4-(((1S,2S)-2-((2,2-difluoroethyl)(ethyl)amino)-cyclopentyl)oxy)benzoate (27-3a)

To a stirred solution of 27-2a (28.2 mg, 0.087 mmol) in EtOH (1 mL) was added SOCl₂ (0.04 mL, 0.55 mmol) and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was then diluted with water (10 mL), neutralized with sat. aq. NaHCO₃, and extracted with EtOAc (3×30 mL). The combined organic phases were then washed with 5% aq. NaHCO₃ (10 mL), brine (10 mL), dried over MgSO₄, filtered, and concentrated to dryness to afford the crude 27-3a as a black gum which was carried onto the next step without further purification (29.7 mg, 0.061 mmol, 70% yield). MS [M+H]⁺=390.4. ¹H NMR (400 MHz, DCM-d₂) δ 8.00 (d, J=8.7 Hz, 1H), 7.16 (d, J=2.6 Hz, 1H), 6.97 (dd, J=8.8, 2.6 Hz, 1H), 5.80 (tt, J=56.4, 4.3 Hz, 1H), 5.08 (s, 2H), 4.63 (dt, J=7.6, 3.5 Hz, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.42 (ddd, J=9.6, 7.5, 4.5 Hz, 1H), 2.98-2.84 (m, 2H), 2.81-2.64 (m, 2H), 2.04 (dddd, J=12.4, 10.8, 6.8, 2.5 Hz, 2H), 1.89-1.74 (m, 2H), 1.61-1.48 (m, 2H), 1.41 (t, J=7.1 Hz, 3H), 1.10 (t, J=7.1 Hz, 3H).

Step 3. 3-(5-(((1S,2S)-2-((2,2-difluoroethyl)(ethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-1)

To a stirred solution of 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 17 mg, 0.10 mmol) and i-Pr₂NEt (0.04 mL, 0.24 mmol) in DMF (0.5 mL) was added a solution of 27-3a (29.7 mg, 0.076 mmol) in DMF (0.5 mL) and the resulting mixture was stirred at 85° C. overnight. The reaction mixture was then cooled to room temperature, diluted with EtOAc (40 mL), washed with sat. aq. NaHCO₃ (3×10 mL), brine (10 mL), dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and lyophilized. The product was repurified by silica gel chromatography eluting with 10% Et₃N in EtOAc to afford the I-1 as a white solid (1.7 mg, 3.79 μmol, 5% yield). MS [M+H]⁺=436.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.21 (d, J=2.1 Hz, 1H), 7.09-7.02 (m, 1H), 6.16-5.78 (m, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.67 (s, 1H), 4.48-4.16 (m, 2H), 2.97-2.78 (m, 2H), 2.64-2.59 (m, 1H), 2.42-2.36 (m, 2H), 2.10-2.03 (m, 3H), 2.01-1.89 (m, 3H), 1.74-1.56 (m, 3H), 1.48 (d, J=10.6 Hz, 1H), 0.99 (t, J=7.0 Hz, 3H).

Example 28: 3-(5-(((1S,2S)-2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-16)

Step 1. 5-(((1S,2S)-2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)isobenzofuran-1(3H)-one (28-2a)

Cyclopentane-1,3-dicarbaldehyde 28-1a was prepared as reported in European Journal of Organic Chemistry, 2007, 1, 53-61

To a stirred solution of 26-2a (186 mg, 0.537 mmol) and NaBH(OAc)₃ (243 mg, 1.15 mmol) in DMF (3 mL) under an atmosphere of nitrogen was added a solution of cyclopentane-1,3-dicarbaldehyde 28-1a (79 mg, 0.63 mmol) in DMF (2.5 mL) and the resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with water (20 mL), basified with sat. aq. NaHCO₃ and extracted with EtOAc (3×30 mL). The combined organic phases were washed with 0.5 M aq. LiCl, sat. aq. NaHCO₃, and brine, dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 20% MeOH in DCM (with 0.1% Et₃N modifier) to afford the 28-2a as a colorless gum (76.1 mg, 0.181 mmol, 34% yield). MS [M+H]⁺=328.6.

Step 2. Ethyl 4-(((1S,2S)-2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-2-(chloromethyl)benzoate (28-3a)

To a stirred solution of 28-2a (76.1 mg, 0.232 mmol) in EtOH (2.5 mL) was added SOCl₂ (0.085 mL, 1.16 mmol) and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was then diluted with water (10 mL), neutralized with sat. aq. NaHCO₃, and extracted with EtOAc (3×30 mL). The combined organic phases were washed with 5% NaHCO₃ solution (10 mL) and brine (10 mL), dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 15% MeOH in DCM (with 0.1% Et₃N modifier) to afford 28-3a as a yellow gum (56.2 mg, 0.122 mmol, 52% yield). MS [M+H]⁺=392.4. ¹H NMR (400 MHz, DCM-d₂) δ 8.00 (d, J=8.7 Hz, 1H), 7.09 (d, J=2.6 Hz, 1H), 7.02 (dd, J=8.7, 2.6 Hz, 1H), 5.08 (s, 2H), 4.72-4.63 (m, 1H), 4.36 (q, J=7.1 Hz, 2H), 2.93 (td, J=8.0, 3.7 Hz, 1H), 2.75-2.61 (m, 2H), 2.31-1.96 (m, 6H), 1.96-1.85 (m, 1H), 1.85-1.46 (m, 9H), 1.41 (t, J=7.1 Hz, 3H).

Step 3. 3-(5-(((1S,2S)-2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-16)

To a stirred solution of 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 29 mg, 0.17 mmol) in DMF (0.7 mL) and i-Pr₂NEt (0.07 mL, 0.4 mmol) was added a solution of 28-3a (56.2 mg, 0.122 mmol) in DMF (0.7 mL) and the resulting mixture was stirred at 85° C. overnight. The reaction mixture was concentrated to dryness and the crude product was purified by silica gel chromatography eluting with EtOAc to afford a mixture of starting materials and product. The mixture was then dissolved in DMF (1 mL) and heated to 120° C. until complete consumption of starting materials was observed by LC-MS. The reaction mixture was concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford I-16 as a white solid (7.3 mg, 0.016 mmol, 11% yield). MS [M+H]⁺=438.5. ¹H NMR (400 MHz, Chloroform-d) δ 7.88 (s, 1H), 7.80 (d, J=8.4 Hz, 1H), 7.13 (d, J=2.8 Hz, 1H), 7.09 (dt, J=8.4, 2.0 Hz, 1H), 5.23 (dd, J=13.2, 5.2 Hz, 1H), 4.62 (dd, J=7.1, 3.4 Hz, 1H), 4.54-4.25 (m, 2H), 3.01-2.77 (m, 4H), 2.62 (dd, J=18.5, 10.9 Hz, 2H), 2.37 (qd, J=13.1, 5.1 Hz, 1H), 2.30-2.14 (m, 4H), 2.11 (s, 1H), 2.07-1.93 (m, 1H), 1.93-1.58 (m, 8H), 1.52-1.44 (m, 1H), 1.37 (d, J=10.9 Hz, 1H).

Example 29: 3-(5-(((1S,2S)-2-morpholinocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-33)

Step 1. 5-(((1S,2S)-2-morpholinocyclopentyl)oxy)isobenzofuran-1(3H)-one (29-1a)

To a stirred solution of 26-2a (145 mg, 0.42 mmol) and 1-bromo-2-(2-bromoethoxy)ethane (29-1, 111 mg, 0.479 mmol) in DMF (2 mL) was added i-Pr₂NEt (0.30 mL, 1.7 mmol) and the resulting mixture was stirred at 80° C. overnight. The reaction mixture was then diluted with EtOAc (80 mL) and washed with 0.5 M aq. LiCl solution (20 mL), sat. aq. NaHCO₃ (20 mL), and brine (20 mL). The organic layer was then dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 20% MeOH in DCM (with 0.1% Et₃N modifier) to afford the 29-1a as a brown solid (35.7 mg, 0.113 mmol, 27% yield). MS [M+H]⁺=304.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.80 (d, J=8.5 Hz, 1H), 7.06 (dd, J=8.5, 2.2 Hz, 1H), 6.97 (s, 1H), 5.26 (s, 2H), 4.70 (s, 1H), 3.68 (s, 4H), 3.03-2.88 (m, 1H), 2.63-2.39 (m, 4H), 2.22-2.00 (m, 2H), 1.81 (d, J=9.7 Hz, 2H), 1.55 (d, J=11.9 Hz, 2H).

Step 2. Ethyl 2-(chloromethyl)-4-(((1S,2S)-2-morpholinocyclopentyl)oxy)benzoate (29-2a)

To a stirred solution of 29-1a (35.7 mg, 0.113 mmol) in EtOH (1 mL) was added SOCl₂ (0.04 mL, 0.5 mmol) and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was diluted with water (10 mL), neutralized with sat. aq. NaHCO₃, and extracted with EtOAc (3×30 mL). The combined organic phases were then washed with 5% aq. NaHCO₃ (10 mL) and brine (10 mL), dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford 29-2a as an amber gum (25.6 mg, 0.068 mmol, 58% yield). MS [M+H]⁺=368.3. ¹H NMR (400 MHz, DCM-d₂) δ 8.00 (d, J=8.7 Hz, 1H), 7.10 (d, J=2.6 Hz, 1H), 6.91 (dd, J=8.7, 2.6 Hz, 1H), 5.08 (d, J=2.5 Hz, 2H), 4.69 (s, 1H), 4.36 (q, J=7.1 Hz, 2H), 3.68 (s, 4H), 2.93 (s, 1H), 2.54 (s, 3H), 2.18-1.96 (m, 2H), 1.87-1.71 (m, 3H), 1.64-1.49 (m, 2H), 1.41 (t, J=7.1 Hz, 3H).

Step 3. 3-(5-(((1S,2S)-2-morpholinocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-33)

To a stirred solution of 3-aminopiperidine-2,6-dione hydrochloride (1-1c, 14 mg, 0.086 mmol) and i-Pr₂NEt (0.030 mL, 0.17 mmol) in DMF (0.7 mL) was added a solution of 29-2a (25.6 mg, 0.068 mmol) in DMF (0.7 mL) and the resulting mixture was stirred at 90° C. overnight. The reaction mixture was diluted with EtOAc (40 mL), washed with sat. aq. NaHCO₃ (2×10 mL) and brine (10 mL), dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing product were combined and lyophilized to afford I-33 as a white solid (3.5 mg, 8.3 μmol, 12% yield). MS [M+H]⁺=414.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.2 Hz, 1H), 7.04 (dd, J=8.4, 1.9 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.74 (d, J=2.9 Hz, 1H), 4.46-4.16 (m, 2H), 3.55 (t, J=4.6 Hz, 4H), 2.99-2.78 (m, 1H), 2.16-2.06 (m, 1H), 2.04-1.83 (m, 4H), 1.77-1.53 (m, 3H), 1.53-1.41 (m, 1H).

Example 30: Enantiomer 1-3-(5-((1S,2S)-2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-44) and Enantiomer 2-3-(5-((1R,2R)-2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-45)

Step 1. Racemic trans-tert-butyl 2-hydroxycyclobutyl)carbamate (30-2a)

To a stirred suspension of racemic trans-2-aminocyclobutanol hydrochloride (30-1a, 500 mg, 4.05 mmol) and di-tert-butyl dicarbonate (971 mg, 4.45 mmol) in THF (15 mL) was added K₂CO₃ (615 mg, 4.45 mmol) and the resulting mixture was stirred vigorously at room temperature for 48 h. The reaction mixture was diluted with brine (5 mL) and extracted with EtOAc (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane. The fractions containing the desired product were combined and concentrated to afford 30-2a (440 mg, 2.35 mmol, 58% yield) as a white solid. ¹H NMR (400 MHz, DCM-d₂) δ 4.82 (s, 1H), 3.95-3.81 (m, 1H), 3.73-3.53 (m, 1H), 2.13-1.90 (m, 2H), 1.58-1.45 (m, 1H), 1.40 (s, 10H), 1.33-1.19 (m, 1H).

Step 2. Racemic tert-butyl (trans-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclobutyl)carbamate (30-3a)

To 5-bromoisobenzofuran-1(3H)-one (19-1a, 300 mg, 1.41 mmol), racemic trans-tert-butyl-3-hydroxycyclopentyl)carbamate (30-2a, 440 mg, 2.35 mmol), NiCl₂(glyme) (15.5 mg, 0.070 mmol), dtbbpy (18.9 mg, 0.070 mmol), and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (15.8 mg, 0.014 mmol) under an atmosphere of nitrogen was added MeCN (4.7 mL) and 2,2,6,6-tetramethylpiperidine (40-2, 286 μl, 1.69 mmol) and the resulting mixture was stirred vigorously for 16 hours under irradiation of blue LED light at room temperature. The reaction mixture was filtered through a short pad of Celite® and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc (with 1% Et₃N modifier) in heptane to afford 30-3a (445 mg, 1.39 mmol, 99% yield) as a white solid. MS [M-tBu+H]⁺=320.3.

Step 3. Racemic 5-(trans-2-aminocyclobutoxy)isobenzofuran-1(3H)-one CF₃CO₂H Salt (30-4a)

To a stirred solution of 30-3a (660 mg, 2.67 mmol) in DCM (7 mL) was added TFA (1.59 mL, 20.7 mmol) and the resulting mixture was stirred at room temperature overnight. The reaction mixture was concentrated to dryness by azeotroping with PhMe (×3) to afford the trifluoroacetate salt of 30-4a (1.97 g, 5.45 mmol, 96% yield) as a brown oil. The crude product was carried onto the next step without further purification. MS [M+H]⁺=220.4. ¹H NMR (400 MHz, DMSO-d₆) δ 7.77 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.3 Hz, 1H), 7.08 (dd, J=8.5, 2.3 Hz, 1H), 5.34 (s, 2H), 4.89-4.81 (m, 1H), 3.96-3.84 (m, 1H), 2.12-2.03 (m, 1H), 1.94-1.85 (m, 1H), 1.60-1.50 (m, 1H), 1.48-1.36 (m, 1H).

Step 4. 5-((1S,2S)-2-(benzylamino)cyclobutoxy)isobenzofuran-1(3H)-one (30-5a) and 5-((1R,2R)-2-(benzylamino)cyclobutoxy)isobenzofuran-1(3H)-one (30-5b)

To a solution of racemic 30-4a (85 mg, 0.388 mmol) in DCM (2 mL) was added benzaldehyde (0.039 mL, 0.39 mmol) and the reaction mixture was stirred at room temperature overnight. NaBH(OAc)₃ (82 mg, 0.39 mmol) was then added and the resulting mixture was stirred at rt for 1 h. The reaction mixture was then quenched with sat. aq. NaHCO₃ and the aqueous phase was extracted with DCM. The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel column chromatography eluting with 0% to 10% MeOH in DCM to afford the desired racemic trans product (48 mg, 0.155 mmol, 40% yield) as a yellow oil. The enantiomers were separated using chiral SFC (Column: 2.1×25.0 cm Chiralpak AD-H; CO₂ co-solvent: MeOH/MeCN (v/v=1:3) with 0.25% isopropylamine; Isocratic method: 25% co-solvent at 80 g/min; System pressure: 125 bar; Column temperature: 25° C.; Sample diluent: MeOH) to afford enantiomer 1 (first peak, Rt=2.70 min, 16.9 mg, 0.055 mmol) and enantiomer 2 (second peak, Rt=3.39 min, 14.9 mg, 0.048 mmol). The absolute stereochemistry of the two enantiomers corresponding to the two product peaks is unknown and was assigned arbitrarily. MS [M+H]⁺=310.3.

Step 5 and 6. Enantiomer 1-3-(5-((1S,2S)-2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-44)

To a stirred solution of enantiomer 1 (30-5a, 16.9 mg, 0.055 mmol) in EtOH (1 mL) was added SOCl₂ (0.04 mL, 0.55 mmol) and the resulting mixture was stirred at 70° C. overnight. The solution was allowed to cool to room temperature and then concentrated to dryness by azeotroping with PhMe (×3) to afford the crude 30-6a as a brown-red gum. MS [M+H]⁺=374.4.

To a stirred solution of 3-aminopiperidine-2,6-dione hydrochloride (1-1c, 9.0 mg, 0.055 mmol) and i-Pr₂NEt (0.05 mL, 0.3 mmol) in DMF (0.5 mL) was added a solution of crude 30-6a (˜0.055 mmol) in DMF (0.5 mL) and the resulting mixture was stirred at 85° C. overnight and then at 110° C. for 5 h. The reaction mixture was concentrated to dryness by azeotroping with PhMe and the resulting crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 10 mM NH₄OH). The purified material was collected into tubes containing several drops of formic acid. The fractions containing product were combined and concentrated by lyophilizer to afford the formate salt of I-44 (enantiomer 1, 3 mg, 6.1 μmol, 11% yield) as a light brown powder. Absolute stereochemistry is unknown and was assigned arbitrarily. MS [M+H]⁺=420.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.38 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.34-7.27 (m, 4H), 7.24-7.17 (m, 2H), 7.06 (dd, J=8.4, 1.0 Hz, 1H), 6.59 (s, 1H), 5.07 (ddd, J=13.3, 5.1, 1.5 Hz, 1H), 4.50 (q, J=7.3 Hz, 1H), 4.38 (dd, J=17.2, 10.5 Hz, 1H), 4.25 (dd, J=17.2, 6.4 Hz, 1H), 3.76-3.66 (m, 2H), 2.91 (ddd, J=18.0, 13.6, 5.4 Hz, 1H), 2.63-2.59 (m, 1H), 2.04-1.94 (m, 2H), 1.50 (quint, J=9.9 Hz, 1H), 1.36 (quint, J=9.5 Hz, 1H).

Step 7 and 8. Enantiomer 2-3-(5-((1R,2R)-2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-45)

To a stirred solution of enantiomer 2 (30-5b, 14.9 mg, 0.048 mmol) in EtOH (1 mL) was added SOCl₂ (0.07 mL, 0.96 mmol) and the resulting reaction mixture was stirred at 70° C. overnight. The solution was allowed to cool to room temperature and concentrated to dryness by azeotroping with PhMe (×3) to afford the crude 30-6b as a brown-red gum. MS [M+H]⁺=374.4.

To a stirred solution of 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 7.9 mg, 0.048 mmol) and i-Pr₂NEt (0.04 mL, 0.2 mmol) in DMF (0.5 mL) was added a solution of crude 30-6b (˜0.048 mmol) in DMF (0.5 mL) and the resulting mixture was stirred at 85° C. overnight and then at 110° C. for 5 h. The reaction was concentrated to dryness by azeotroping with PhMe and the resulting crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 10 mM NH₄OH). The purified material was collected into tubes containing several drops of formic acid. Fractions containing product were combined and concentrated by lyophilizer to afford I-45 (enantiomer 2, 3 mg, 6 μmol, 13% yield) as a light brown powder. Absolute stereochemistry is unknown and was assigned arbitrarily. MS [M+H]⁺=420.4. ¹H NMR (400 MHz, MeCN-d₃) δ 7.52 (d, J=8.4 Hz, 1H), 7.28-7.21 (m, 2H), 7.19-7.13 (m, 2H), 7.12-7.05 (m, 2H), 6.98 (dd, J=8.4, 2.2 Hz, 1H), 5.03 (ddd, J=13.3, 5.2, 1.2 Hz, 1H), 4.43 (q, J=6.8 Hz, 1H), 4.36-4.23 (m, 2H), 3.75-3.64 (m, 2H), 3.31-3.22 (m, 1H), 2.64 (ddd, J=17.5, 4.5, 2.4 Hz, 1H), 2.47-2.33 (m, 1H), 2.30-2.19 (m, 1H), 2.09-1.99 (m, 2H), 1.49 (tdd, J=10.8, 9.4, 7.9 Hz, 1H), 1.37-1.27 (m, 1H).

Example 31: 3-(5-(((3aR*,6S*,6aS*)-1-methyloctahydrocyclopenta[b]pyrrol-6-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-3)

Step 1. Racemic tert-butyl (3aR*,6S*,6aS*)-6-hydroxyhexahydrocyclopenta[b]pyrrole-1(2H)-carboxylate (31-2a)

To a stirred solution of racemic (3aR*,6S*,6aS*)-octahydrocyclopenta[b]pyrrol-6-ol hydrochloride (31-1a, 1.00 g, 6.11 mmol) and di-tert-butyl dicarbonate (1.60 g, 7.33 mmol) in MeOH (10 mL) was added Na₂CO₃ (0.777 g, 7.33 mmol) and the resulting mixture was stirred vigorously at room temperature for 48 hours. The reaction mixture was then diluted with brine (25 mL) and extracted with DCM (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness to afford 31-2a (1.32 g, 5.81 mmol, 95% yield) as a viscous yellow oil. The product was carried onto the next step without further purification. ¹H NMR (400 MHz, DCM-d₂) δ 3.95 (dt, J=9.0, 4.9 Hz, 1H), 3.74 (dd, J=9.1, 4.3 Hz, 1H), 3.42-3.28 (m, 2H), 2.87-2.73 (m, 1H), 1.98-1.87 (m, 3H), 1.64-1.48 (m, 3H), 1.44 (s, 9H), 1.37-1.27 (m, 1H).

Step 2. 3-(5-bromo-1-oxoisoindolin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (31-3a)

To a stirred solution of 1-1d (10.0 g, 30.9 mmol) and DBU (6.9 mL, 46 mmol) in DMF (95 mL) was added SEMCl (6.6 mL, 37 mmol) at 0° C. and the resulting reaction mixture was allowed to warm to room temperature and then stirred for 5 h. An additional portion of DBU (3.5 mL, 23 mmol) and SEMCl (3.3 mL, 19 mmol) was added and the reaction mixture was stirred for additional 2 h. The reaction mixture was then quenched with sat. aq. NH₄Cl (250 mL) and extracted with EtOAc (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was dissolved in minimal amount of EtOAc (˜50 mL) and Et₂O:heptane (v/v=1:2, 400 mL) was added. The resulting cloudy solution was left standing at −5° C. overnight. The formed precipitate was filtered, washed with heptane (×3), and dried under vacuum to afford 31-3a (11.53 g, 25.4 mmol, 82% yield) as an off-white solid. MS [M+H]⁺=453.4. ¹H NMR (400 MHz, Chloroform-d) δ 7.75 (d, J=8.6 Hz, 1H), 7.66-7.61 (m, 2H), 5.37-5.09 (m, 3H), 4.48 (d, J=16.2 Hz, 1H), 4.32 (d, J=16.2 Hz, 1H), 3.74-3.50 (m, 2H), 3.11-2.98 (m, 1H), 2.94-2.83 (m, 1H), 2.33 (qd, J=13.2, 4.7 Hz, 1H), 2.24-2.15 (m, 1H), 0.97-0.90 (m, 2H), 0.00 (s, 9H).

Step 3. Racemic tert-butyl (3aR*,6S*,6aS*)-6-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)-methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)hexahydrocyclopenta[b]pyrrole-1 (2H)-carboxylate (31-4a)

To a stirred solution of 31-3a (1396 mg, 3.08 mmol), 31-2a (700 mg, 3.08 mmol), NiCl₂(glyme) (34 mg, 0.15 mmol), dtbbpy (41 mg, 0.15 mmol) and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (34.5 mg, 0.031 mmol) in MeCN (5 mL) under an atmosphere of nitrogen was added 2,2,6,6-tetramethylpiperidine (40-2, 0.52 mL, 3.1 mmol) and the resulting mixture was stirred vigorously overnight under irradiation of blue LED light at room temperature. The reaction mixture was then diluted with DCM (20 mL), filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford 31-4a (840 mg, 1.40 mmol, 45.5% yield) as a yellow solid. MS [M−H]⁻=598.6. ¹H NMR (400 MHz, DCM-d₂) δ 7.67 (d, J=8.4 Hz, 1H), 7.47-6.91 (m, 2H), 5.23-5.06 (m, 3H), 5.00-4.76 (m, 1H), 4.37-4.22 (m, 2H), 4.14-4.09 (m, 1H), 3.65-3.49 (m, 3H), 3.17 (br s, 1H), 3.01-2.79 (m, 3H), 2.37-2.01 (m, 3H), 2.00-1.68 (m, 3H), 1.65-1.57 (m, 1H), 1.45-1.35 (m, 10H), 0.93-0.85 (m, 2H), −0.02 (s, 9H).

Step 4 and 5. Racemic 3-(5-(((3aR*,6S*,6aS*)-1-methyloctahydrocyclopenta[b]pyrrol-6-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-3)

To a stirred solution of 31-4a (210 mg, 0.350 mmol) in DCM (1 mL) was added TFA (1 mL, 13.07 mmol) and the resulting reaction mixture stirred for 2 h at room temperature. The reaction mixture was then concentrated to dryness by azeotroping with PhMe (×3) to afford a mixture of 31-5a and 31-5b based on a crude ¹H NMR analysis. The crude mixture was carried onto the next step without further purification.

To a crude mixture of 31-5a and 31-5b (˜0.35 mmol) in DMF (1 mL) was added acetaldehyde (2-1, 0.06 mL, 1.1 mmol), followed by NaBH(OAc)₃ (148 mg, 0.700 mmol) and the resulting mixture was stirred for 6 h at room temperature. LC-MS analysis indicated formation of I-3 (MS [M+H]⁺=384.1, formed via reductive amination with formaldehyde that was released by 31-5b and 31-6b (MS [M+H]⁺=398.3) in a 2:1 ratio, respectively. The reaction mixture was then concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 5 mM NH₄OH) and collected into tubes containing formic acid (2 drops). The fractions containing the desired product were combined and lyophilized to afford the formate salt of racemic I-3 (10.1 mg, 0.023 mmol, 7% yield). MS of I-3 [M+H]⁺=384.1. ¹H NMR (400 MHz, DCM-d₂) δ 8.26 (s, 1H), 7.80-7.62 (m, 1H), 7.08-6.92 (m, 2H), 5.11 (dd, J=13.3, 5.1 Hz, 1H), 4.63 (s, 1H), 4.38-4.23 (m, 2H), 3.00-2.70 (m, 5H), 2.53 (s, 1H), 2.42 (s, 3H), 2.38-2.12 (m, 3H), 2.04-1.93 (m, 3H), 1.89-1.81 (m, 1H), 1.53-1.35 (m, 2H).

Example 32: 3-(5-(((3R,4S)-4-(diethylamino)tetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-47)

Step 1. tert-butyl ((3S,4R)-4-hydroxytetrahydrofuran-3-yl)carbamate (32-2a)

To a stirred solution of (3R,4S)-4-aminotetrahydrofuran-3-ol (32-1a, 482 mg, 4.67 mmol) and di-tert-butyl dicarbonate (1122 mg, 5.14 mmol) in MeOH (10 mL) was added Et₃N (1.6 mL, 11.48 mmol) and the resulting mixture was stirred at room temperature until complete consumption of starting material was observed. The reaction mixture was then concentrated to dryness. The crude material was treated with water (6 mL) and the resulting solid was then filtered off and washed with water to afford the desired product 32-2a (838.1 mg, 4.08 mmol, 87% yield) as a white solid. MS [M-tBu+H]⁺=148.2. ¹H NMR (400 MHz, Chloroform-d) δ 4.71 (s, 1H), 4.32 (dq, J=5.7, 2.9 Hz, 1H), 4.10 (ddd, J=16.0, 9.7, 5.6 Hz, 2H), 3.97 (dt, J=6.4, 3.2 Hz, 1H), 3.71 (dd, J=10.0, 3.3 Hz, 1H), 3.63 (dd, J=9.5, 3.4 Hz, 1H), 2.82 (s, 1H), 1.48 (s, 9H).

Step 2. tert-butyl ((3S,4R)-4-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)tetrahydrofuran-3-yl)carbamate (32-3a)

To a stirred solution of (31-3a, 934.6 mg, 2.061 mmol), tert-butyl ((3S,4R)-4-hydroxytetrahydrofuran-3-yl)carbamate (32-2a, 395.4 mg, 1.946 mmol), NiCl₂(glyme) (25 mg, 0.11 mmol), dtbbpy (30 mg, 0.11 mmol), and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (20 mg, 0.018 mmol) in MeCN (5 mL) under an atmosphere of nitrogen was added 2,2,6,6-tetramethylpiperidine (40-2, 0.35 mL, 2.1 mmol) and the resulting mixture was stirred overnight under irradiation of blue LED light at room temperature. The reaction mixture was diluted with EtOAc and filtered through a pad of Celite®. The filtrate was then diluted with EtOAc (150 mL), washed with water (30 mL) and brine (20 mL), dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptane to afford 32-3a as a white solid (1.02 g, 1.59 mmol, 82% yield). MS [M+18]⁺=593.6. ¹H NMR (400 MHz, DMSO-d₆) δ 7.66 (d, J=8.4 Hz, 1H), 7.43 (d, J=6.3 Hz, 1H), 7.26 (s, 1H), 7.11 (dd, J=8.5, 2.2 Hz, 1H), 5.30-5.00 (m, 3H), 4.88 (d, J=4.2 Hz, 1H), 4.52-4.20 (m, 2H), 4.05 (qd, J=6.2, 5.2, 3.4 Hz, 4H), 3.84 (d, J=10.3 Hz, 1H), 3.64-3.50 (m, 3H), 3.08 (ddd, J=18.4, 13.6, 5.5 Hz, 1H), 2.82 (d, J=4.3 Hz, 1H), 2.46-2.36 (m, 1H), 2.10-2.02 (m, 1H), 1.43 (s, 9H), 1.20 (t, J=7.1 Hz, 1H), 0.94-0.75 (m, 2H). Note: TMS peak of SEM group is overlapping with reference Me₄Si.

Step 3. 3-(5-(((3R,4S)-4-aminotetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-46)

To a stirred solution of 32-3a (102 mg, 0.177 mmol) in MeCN (1 mL) was added 1 M HCl in acetic acid (0.9 mL, 0.9 mmol) and the resulting mixture was stirred at room temperature until complete consumption of starting materials was observed. The reaction mixture was then concentrated to dryness by azeotroping with DCM to afford the crude product I-46 as a white foam which was carried onto the next step without further purification. MS [M+H]⁺=346.4. ¹H NMR (400 MHz, DMSO-d₆) δ 11.96 (s, 1H), 8.38 (s, 3H), 7.70 (dd, J=8.4, 1.1 Hz, 1H), 7.27 (dd, J=7.4, 2.3 Hz, 1H), 7.17-7.09 (m, 1H), 5.30-4.94 (m, 2H), 4.54-4.20 (m, 2H), 4.08 (dd, J=10.2, 5.5 Hz, 1H), 3.93 (s, 1H), 3.87-3.76 (m, 2H), 3.05 (ddd, J=18.2, 13.5, 5.4 Hz, 1H), 2.88-2.71 (m, 1H), 2.43-2.34 (m, 1H), 2.07-2.00 (m, 1H).

Step 4. 3-(5-(((3R,4S)-4-(diethylamino)tetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-47)

To a solution of crude I-46 (65 mg, ˜0.17 mmol) and acetaldehyde (2-1, 0.03 mL, 0.5 mmol) in DMF (1 mL) was added NaBH(OAc)₃ (108 mg, 0.511 mmol) in one portion and the resulting mixture was stirred at room temperature open to air for 4 hours. An additional portion of acetaldehyde (2-1, 0.03 mL, 0.5 mmol) and NaBH(OAc)₃ (69 mg, 0.32 mmol) were then added and the reaction mixture was stirred at room temperature until complete consumption of starting material was observed. The reaction mixture was then concentrated to dryness. The crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 5 mM NH₄OH) and collected into tubes containing formic acid (2 drops). The fractions containing the desired product were combined and lyophilized to afford the formate salt of I-47 as a white solid (39.7 mg, 0.087 mmol, 51% yield). MS [M+H]⁺=402.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.23 (s, 1H), 7.63 (d, J=8.4 Hz, 1H), 7.25 (d, J=2.1 Hz, 1H), 7.09 (ddd, J=8.5, 2.3, 1.1 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.90 (t, J=3.7 Hz, 1H), 4.48-4.19 (m, 2H), 4.08 (dd, J=10.5, 5.4 Hz, 1H), 3.99 (dd, J=9.2, 7.0 Hz, 1H), 3.72 (dt, J=10.4, 2.0 Hz, 1H), 3.60 (dd, J=9.2, 6.5 Hz, 1H), 3.47 (td, J=6.8, 2.5 Hz, 1H), 2.91 (ddd, J=17.3, 13.7, 5.5 Hz, 1H), 2.56 (t, J=7.0 Hz, 4H), 2.39 (dd, J=13.2, 4.5 Hz, 1H), 2.05-1.91 (m, 1H), 0.97 (t, J=7.0 Hz, 6H).

Example 33: 3-(5-(((1S,2S)-2-methoxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-4)

To a suspension of 1-1d (200 mg, 0.619 mmol) and (1S,2S)-2-methoxycyclohexan-1-ol (33-1a, 121 mg, 0.928 mmol) in MeCN (5 mL), whilst bubbling with nitrogen, was added Ir[dF(CF₃)ppy)₂dtbbpy)]PF₆ (7 mg, 6 μmol), NiCl₂(glyme) (7 mg, 0.03 mmol), dtbbpy (8 mg, 0.03 mmol), quinuclidine (7 mg, 0.06 mmol) and K₂CO₃ (128 mg, 0.928 mmol). The resulting suspension was sonicated and stirred vigorously under irradiation of blue LED light at room temperature for 16 h. The reaction mixture was diluted with excess EtOAc and the resulting solid was filtered and rinsed with EtOAc (×3). The filtrate was concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0% to 25% EtOH:EtOAc (v/v=1:3) in DCM, evaporating the appropriate fractions to dryness. Isolated material was purified by reverse phase HPLC (MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and concentrated by lyophilizer to afford I-4 (7.0 mg, 0.018 mmol, 3% yield) as a white solid. MS [M+H]⁺=373.3. ¹H NMR (400 MHz, DMSO-d₆): δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.20 (s, 1H), 7.07 (d, J=8.9 Hz, 1H), 5.06 (dd, J=13.1, 5.0 Hz, 1H), 4.41-4.21 (m, 3H), shoulder on H₂O signal at 3.30 (3H), 2.96-2.84 (m, 1H), 2.64-2.56 (m, 3H), 2.43-2.34 (m, 1H), 2.07-1.94 (m, 2H), 1.69-1.57 (m, 2H), 1.45-1.22 (m, 4H)

Example 34: 3-(5-(((1S,2S)-2-methoxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-26)

To a suspension of 1-1d (200 mg, 0.619 mmol) and (1S,2S)-2-methoxycyclopentan-1-ol (34-1a, 108 mg, 0.928 mmol) in MeCN (5 mL), whilst bubbling with nitrogen, was added Ir[dF(CF₃)ppy)₂dtbbpy)]PF₆ (7 mg, 6 μmol), NiCl₂(glyme) (7 mg, 0.03 mmol), dtbbpy (8 mg, 0.03 mmol), quinuclidine (7 mg, 0.06 mmol) and K₂CO₃ (128 mg, 0.928 mmol). The resulting suspension was sonicated and stirred vigorously under irradiation of blue LED light at room temperature for 16 h. The reaction mixture was diluted with excess EtOAc and the resulting solid was filtered and rinsed with EtOAc (×3). The filtrate was concentrated to dryness and the crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and concentrated by lyophilizer to afford I-26 (38 mg, 0.10 mmol, 16% yield) as a white solid. MS [M+H]⁺=359.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.17 (d, J=2.1 Hz, 1H), 7.05 (dd, J=8.5, 2.1 Hz, 1H), 5.07 (dd, J=13.3, 5.2 Hz, 1H), 4.76-4.71 (m, 1H), 4.39 (dd, J=17.2, 4.5 Hz, 1H), 4.26 (dd, J=17.3, 3.3 Hz, 1H), 3.84-3.78 (m, 1H), 3.28 (s, 3H), 2.98-2.83 (m, 1H), 2.63-2.54 (m, 1H), 2.43-2.33 (m, 1H), 2.16-2.06 (m, 1H), 2.03-1.88 (m, 2H), 1.74-1.58 (m, 4H).

Example 35: 3-(1-oxo-5-(((1S,2S)-2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione and 3-(1-oxo-5-(((1R,2R)-2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-19)

Step 1. (1S,2S)-2-phenoxycyclohexan-1-ol and (1R,2R)-2-phenoxycyclohexan-1-ol (35-2a)

Cyclohexene oxide (35-1a, 1.96 g, 20 mmol), phenol (0.94 g, 10 mmol) and tetra-n-butyl ammonium iodide (0.37 g, 1 mmol) were stirred at 80° C. for 70 h. The reaction mixture was then diluted with EtOAc (50 mL) and washed with 10% aq. NaHCO₃ (3×20 mL) and water (2×15 mL). The organic layer was dried over MgSO₄, filtered, and concentrated to dryness. The crude material was purified using silica gel chromatography eluting with PhMe:EtOAc (v/v=10:1) to afford 35-2a (1.76 g, 8.98 mmol, 90% yield) as a white solid. MS [M+Na]⁺=215.1. ¹H NMR (400 MHz, Chloroform-d) δ 7.27-7.22 (m, 2H), 6.98-6.91 (m, 3H), 4.05-3.96 (m, 1H), 3.88-3.84 (m, 1H), 2.54 (br s, 1H), 2.20-2.06 (m, 2H), 1.79-1.71 (m, 2H), 1.43-1.22 (m, 4H).

Step 2. 3-(1-oxo-5-(((1S,2S)-2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione and 3-(1-oxo-5-(((1R,2R)-2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-19)

To a suspension of 1-1d (200 mg, 0.619 mmol) and 2-phenoxycyclohexan-1-ol (35-2a, mixture of trans isomers) (178 mg, 0.928 mmol) in MeCN (5 mL), whilst bubbling with nitrogen, was added Ir[dF(CF₃)ppy)₂dtbbpy)]PF₆ (7 mg, 6 μmol), NiCl₂(glyme) (7 mg, 0.03 mmol), dtbbpy (8 mg, 0.03 mmol), quinuclidine (7 mg, 0.06 mmol) and K₂CO₃ (128 mg, 0.928 mmol). The resulting suspension was sonicated and then stirred vigorously under irradiation of blue LED light at room temperature for 3 days. The reaction mixture was diluted with excess EtOAc and the resulting solid was filtered and rinsed with EtOAc (3×). The filtrate was concentrated to dryness and the crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and concentrated by lyophilizer to afford I-19 (6.0 mg, 0.013 mmol, 2% yield) as a white solid. MS [M+H]⁺=435.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.58 (d, J=8.5 Hz, 1H), 7.24 (t, J=7.4 Hz, 2H), 7.19 (s, 1H), 7.05-7.01 (m, 1H), 6.95-6.87 (m, 3H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.62-4.55 (m, 1H), 4.51-4.42 (m, 1H), 4.42-4.18 (m, 2H), 2.90 (m, 1H), 2.64-2.54 (m, 2H), 2.42-2.32 (m, 1H), 2.19-2.10 (m, 1H), 2.03-1.92 (m, 1H), 1.75-1.64 (m, 2H), 1.58-1.38 (m, 4H).

Example 36: 3-(5-(((1S,2S)-2-(benzyloxy)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-9)

To a suspension of 1-1d (200 mg, 0.619 mmol) and (1S,2S)-2-(benzyloxy)cyclopentan-1-ol (36-1a, 268 mg, 1.39 mmol) in MeCN (5 mL), whilst bubbling with nitrogen, was added Ir[dF(CF₃)ppy)₂dtbbpy)]PF₆ (7 mg, 6 μmol), NiCl₂(glyme) (7 mg, 0.03 mmol), dtbbpy (8 mg, 0.03 mmol), quinuclidine (7 mg, 0.06 mmol) and K₂CO₃ (128 mg, 0.928 mmol). The resulting suspension was sonicated and then stirred vigorously under irradiation of blue LED light at room temperature for 3 days. The reaction mixture was diluted with excess EtOAc and the resulting solid was filtered and rinsed with EtOAc (×3). The filtrate was concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0% to 20% THF in DCM. The fractions containing the desired product were concentrated and further purified by reverse phase HPLC (eluting with MeCN/H₂O containing 0.1% formic acid). The fractions containing the desired product were combined and concentrated by lyophilizer to afford I-9 (21 mg, 0.046 mmol, 5% yield) as a white solid. MS [M+H]⁺=435.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.37-7.32 (m, 4H), 7.31-7.25 (m, 1H), 7.15 (s, 1H), 7.05-7.01 (m, 1H), 5.12-5.02 (m, 1H), 4.83-4.79 (m, 1H), 4.54 (qd, J=12.0, 1.6 Hz, 2H), 4.43-4.30 (m, 1H), 4.25 (dd, J=17.2, 8.7 Hz, 1H), 4.02-3.98 (m, 1H), 2.96-2.85 (m, 1H), 2.64-2.54 (m, 1H), 2.43-2.34 (m, 1H), 2.20-2.12 (m, 1H), 2.02-1.91 (m, 2H), 1.78-1.64 (m, 4H).

Example 37: 3-(5-(((1R,2R)-2-hydroxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-6)

To a suspension of 1-1d (200 mg, 0.619 mmol) and ((1R,2R)-cyclopentane-1,2-diol (37-1a, 95 mg, 0.93 mmol) in MeCN (5 mL), whilst bubbling with nitrogen, was added Ir[dF(CF₃)ppy)₂dtbbpy)]PF₆ (7 mg, 6 μmol), NiCl₂(glyme) (7 mg, 0.03 mmol), dtbbpy (8 mg, 0.03 mmol), quinuclidine (7 mg, 0.06 mmol) and K₂CO₃ (128 mg, 0.928 mmol). The resulting suspension was sonicated and then stirred vigorously under irradiation of blue LED light at room temperature for 3 days. The reaction mixture was diluted with excess EtOAc and the resulting solid was filtered and rinsed with EtOAc (×3). The filtrate was concentrated to dryness and the crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and concentrated by lyophilizer to afford I-6 (17 mg, 0.047 mmol, 8% yield) as a white solid. MS [M+H]⁺=345.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.16 (d, J=1.7 Hz, 1H), 7.04 (dd, J=8.3, 2.01 Hz, 1H), 5.06 (dd, J=13.5, 5.1 Hz, 1H), 5.01 (t, J=4.0 Hz, 1H), 4.58-4.52 (m, 1H), 4.39 (dd, J=17.2, 5.1 Hz, 1H), 4.26 (dd, J=17.0, 3.9 Hz, 1H), 4.11-4.05 (m, 1H), 2.96-2.85 (m, 1H), 2.64-2.51 (m, 1H), 2.44-2.33 (m, 1H), 2.20-2.09 (m, 1H), 2.02-1.94 (m, 1H), 1.92-1.82 (m, 1H), 1.81-1.50 (m, 4H).

Example 38. 3-(5-(((1S,2R)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-2)

Step 1. tert-butyl ((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)carbamate (38-2a)

To a suspension of 1-Id (432 mg, 1.34 mmol)), tert-butyl ((1S,2S)-2-hydroxycyclohexyl) carbamate (38-1a, 1066 mg, 4.95 mmol), NiCl₂(glyme) (54 mg, 0.25 mmol), dtbbpy (66 mg, 0.2548 mmol), Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (15 mg, 0.013 mmol) in MeCN (4.5 mL) under an atmosphere of nitrogen was added 2,2,6,6-tetramethylpiperidine (40-2, 226 μL, 1.34 mmol) and the resulting mixture was then stirred vigorously for 16 h under irradiation of blue LED light at room temperature. The reaction mixture was then diluted with EtOAc (20 mL), filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0% to 100% EtOAc in heptanes to afford 38-2a (115 mg, 0.251 mmol, 19% yield) as a white powder. MS [M+H]⁺=458.3.

Step 2. 3-(5-(((1S,2R)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione CF₃CO₂H Salt (I-30)

To a stirred solution of 38-2a (115 mg, 0.251 mmol) in DCM (2.5 mL) was added TFA (194 μL, 2.51 mmol) and the resulting mixture was stirred for 48 h. The reaction mixture was concentrated to dryness and the crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid) to afford the trifluoroacetate salt of I-30 (54 mg, 0.152 mmol, 60% yield). MS [M+H]⁺=358.4. ¹H NMR (400 MHz, DMSO-de) δ 10.97 (s, 1H), 7.68 (d, J=8.4 Hz, 1H), 7.27 (d, J=2.0 Hz, 1H), 7.17 (dt, J=8.4, 1.9 Hz, 1H), 5.08 (dd, J=13.3, 5.1 Hz, 1H), 4.76 (s, 1H), 4.41 (dd, J=17.4, 2.4 Hz, 1H), 4.28 (dd, J=17.3, 4.2 Hz, 1H), 3.42 (d, J=6.9 Hz, 1H), 2.91 (ddd, J=18.0, 13.6, 5.4 Hz, 1H), 2.06-1.93 (m, 2H), 1.75 (d, J=23.0 Hz, 3H), 1.64-1.49 (m, 1H), 1.40 (s, 3H).

Step 3. 3-(5-(((1S,2R)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-2)

A solution of I-30 (54.2 mg, 0.152 mmol) and benzaldehyde (15 μL, 0.15 mmol) in DMF (0.7 mL) was stirred for 3 h. NaBH(OAc)₃ (31 mg, 0.15 mmol) was then added in one portion and the resulting reaction mixture was stirred vigorously for 16 h at room temperature. The reaction mixture was concentrated to dryness and the resulting crude material was purified by reverse phase HPLC (eluting with MeCN/H₂O with 0.1% formic acid). The fractions containing the desired product were combined and lyophilized to afford to afford I-2 (13.2 mg, 0.029 mmol, 20% yield) as a white solid. MS [M+H]⁺=448.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.28-7.14 (m, 4H), 7.10 (d, J=8.3 Hz, 1H), 6.51 (s, 1H), 5.07 (dd, J=13.2, 5.1 Hz, 1H), 4.73 (s, 1H), 4.37 (dd, J=17.1, 6.7 Hz, 1H), 4.25 (dd, J=17.5, 4.8 Hz, 1H), 3.72 (q, J=13.1 Hz, 2H), 2.97-2.85 (m, 1H), 2.76 (s, 1H), 1.99 (s, 2H), 1.91 (s, 1H), 1.66 (s, 3H), 1.49 (s, 2H), 1.42-1.21 (m, 1H).

Example 39: 3-(5-(((1S,2S)-2-(3-ethoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-48)

Method A: Step 1. 1,3-dibromo-2-ethoxypropane (39-2)

To a stirred solution of N1,N1,N8,N8-tetramethylnaphthalene-1,8-diamine (80.0 g, 372 mmol) and 1,3-dibromopropan-2-ol (39-1, 20.0 mL, 196 mmol) in DCM (150 mL) was added dropwise triethyloxonium tetrafluoroborate (63.3 g, 333 mmol), dissolved in DCM (50 mL) at 0° C. under positive pressure of nitrogen. After the addition was complete the reaction mixture was stirred at rt for 18 h. The mixture was filtered through a pad of silica gel eluting with DCM (5×350 mL). Each fraction was collected separately and the fractions containing product were combined and concentrated. The yellow oil was purified by filtration through a second silica gel pad eluting with 20% DCM in pentane to afford 39-2 (43.4 g, 159 mmol, 81% yield) was obtained as a colorless oil. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 3.72-3.66 (m, 1H), 3.62 (q, J=7.0 Hz, 2H), 3.55 (d, J=5.2 Hz, 4H), 1.21 (t, J=7.0 Hz, 3H).

Step 2. 3-(5-(((1S,2S)-2-(3-ethoxyazetidin-1-ylcyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-48)

To a stirred suspension of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 7.10 g, 17.9 mmol) and DIPEA (12.5 mL, 71.5 mmol) in DMA (100 mL) was added 39-2 (8.31 g, 30.4 mmol) and the resulting mixture was heated at 85° C. for 24 h under atmosphere of nitrogen. Additional 39-1 (4.0 g, 15 mmol) and DIPEA (6.0 mL, 34 mmol) were added and stirring was continued at 85° C. for an additional 24 h. Further 39-2 (0.80 g, 2.9 mmol) and DIPEA (1.2 mL, 6.9 mmol) were added and stirring was continued at 85° C. for an additional 24 h. The reaction mixture was allowed to cool rt and concentrated to dryness. The crude product was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-48 (3.37 g, 7.63 mmol, 43% yield) as an off-white solid. MS [M+H]⁺=442.5. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 8.39-8.20 (m, 1H), 7.67 (dd, J=8.7, 1.7 Hz, 1H), 7.02-6.93 (m, 2H), 5.09 (ddd, J=13.3, 6.7, 5.2 Hz, 1H), 4.48-4.14 (m, 3H), 4.06-3.87 (m, 1H), 3.59 (s, 2H), 3.34 (q, J=7.0 Hz, 2H), 3.14 (s, 1H), 3.03-2.71 (m, 3H), 2.50-2.22 (m, 2H), 2.20-2.11 (m, 1H), 2.09-1.95 (m, 1H), 1.91-1.80 (m, 1H), 1.76-1.62 (m, 2H), 1.45-1.14 (m, 4H), 1.10 (t, J=7.0 Hz, 3H).

Method B:

Intermediate 54-1 was prepared according to literature procedure described in Bioorganic and Medicinal Chemistry Letters, 2003, 13, 1729-1732.

To a suspension of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 100 mg, 0.28 mmol) in MeCN (3 mL), was added 2-ethoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) (54-1, 180 mg, 0.42 mmol) and DIPEA (0.36 mL, 1.7 mmol) and the resulting mixture was stirred at 120° C. for 16 h under microwave irradiation. The reaction mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over anhydrous Na₂SO₄ and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 6% MeOH in DCM to afford I-48 (20 mg, 0.045 mmol, 16%) as an off-white solid. The analytical data was identical to the one reported herein above for Example 39, Step 2.

Example 40: 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Int-40-5)

Step 1. Tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cycloheptyl)carbamate (40-3)

To a suspension of 3-(5-bromo-1-oxoisoindolin-2-yl)-1-((2-trimethylsilyl)ethoxy)methyl) piperidine-2,6-dione (31-3a) (5.00 g, 11.0 mmol) in MeCN (35 mL), whilst bubbling with nitrogen, was added tert-butyl ((1S,2S)-2-hydroxycycloheptyl)carbamate (40-1, 2.66 g, 11.6 mmol), 2,2,6,6-tetramethylpiperidine (40-2, 1.95 mL, 11.6 mmol), NiCl₂(glyme) (0.121 g, 0.551 mmol), dtbbpy (0.148 g, 0.551 mmol) and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (0.124 g, 0.110 mmol). The suspension was sonicated and then stirred at rt under an atmosphere of nitrogen. The resulting solution was stirred under blue LED lights for 48 hours. A 50% saturated sodium hydrogen carbonate in water was then added and the resulting mixture was extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column, concentrated under reduced pressure and then dried under high vacuum to afford crude 40-3, as a dark red solid, which was carried onto the next step without purification. MS [M−H]⁻=600.5.

Step 2. 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5)

To a solution of crude tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl) piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cycloheptyl)carbamate (40-3, 6.64 g, 11.0 mmol) in DCM (80 mL) under an atmosphere of nitrogen was added methanesulfonic acid (2.9 mL, 44 mmol) and the resulting mixture was stirred at rt overnight. Additional methanesulfonic acid (2.9 mL, 44 mmol) was added and the reaction mixture was stirred at rt for 4 hours and then cooled using an ice bath. Triethylamine (18.5 mL, 132 mmol) was added dropwise, under an atmosphere of nitrogen followed by N1,N2-dimethylethane-1,2-diamine (40-4, 1.4 mL, 13 mmol). The resulting mixture was stirred at rt overnight and then concentrated under reduced pressure. To the resulting residue was added to a 50% saturated sodium hydrogen carbonate in water solution (250 mL). The aqueous phase was extracted with 20% i-PrOH in DCM (×4). The combined organics were passed through a phase separating column and concentrated under reduced pressure. The crude material was treated with MeCN to provide a suspension, which was sonicated and filtered. The collected solid was washed with MeCN (×3) and then dried under vacuum to afford INT-40-5 (2.17 g, 5.84 mmol, 53% yield) as a cream-colored solid. MS [M+H]⁺=372.3. Alternatively, the crude material was treated with excess diethyl ether to provide a suspension, which was sonicated and filtered. The collected solid was washed with diethyl ether (×3) and then was dried under vacuum to afford INT-40-5 (1.42 g, 3.82 mmol, 35% yield) as a pale yellow solid. MS [M+H]⁺=372.3.

Example 41: 3-(5-(((1S,2S)-2-(ethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-57)

To a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 92 mg, 0.25 mmol) in TFE (3 mL) was added acetaldehyde (2-1, 0.02 mL, 0.4 mmol). The resulting mixture was stirred at room temperature for 30 minutes and then sodium triacetoxyborohydride (157 mg, 0.743 mmol) was added. Stirring was continued at room temperature for 3 hours and then a saturated aqueous solution of sodium hydrogen carbonate (20 mL) was added. The reaction mixture was extracted with 20% i-PrOH in DCM (×3) and the combined organic phases were passed through a phase separating column and concentrated under reduced pressure. The crude product was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃), to afford I-57 (64 mg, 0.15 mmol, 61% yield), as a pale yellow solid. MS [M+H]⁺=400.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.14 (s, 1H), 7.03 (dd, J=8.4, 2.2 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.44-4.20 (m, 3H), 2.97-2.82 (m, 2H), 2.70-2.53 (m, 2H), 2.44-2.29 (m, 1H), 2.03-1.92 (m, 1H), 1.84-1.73 (m, 3H), 1.72-1.39 (m, 8H), 0.99 (t, J=7.1 Hz, 3H).

Example 42: 3-(5-(((1S,2S)-2-(benzylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-58)

To a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 46 mg, 0.12 mmol) in MeCN (2 mL) was added DIPEA (0.07 mL, 0.4 mmol) and benzyl bromide (42-1, 0.015 mL, 0.12 mmol). The resulting mixture was stirred at 65° C. overnight. Stirring was continued and then a saturated aqueous solution of sodium hydrogen carbonate (20 mL) was added. The reaction mixture was extracted with 20% i-PrOH in DCM (×3) and the combined organic phases were passed through a phase separating column and concentrated under reduced pressure. The crude product was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N). The resulting material was dissolved in a minimal amount of DCM to which an excess of diethyl ether was added. The resulting suspension was sonicated, filtered, and rinsed with diethyl ether (×3). The obtained filtrate was collected and concentrated. The crude material was purified further via silica gel chromatography eluting with 0-20% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-58 (11 mg, 0.02 mmol, 18% yield), as a white solid. MS [M+H]⁺=462.6. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm); 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.33-7.24 (m, 4H), 7.23-1.17 (m, 1H), 7.11 (s, 1H), 7.00 (dd, J=8.4, 2.1 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.48-4.20 (m, 3H), 3.85-3.63 (m, 2H), 2.96-2.82 (m, 2H), 2.59 (d, J=17.3 Hz, 1H), 2.38 (td, J=13.1, 4.5 Hz, 1H), 2.02-1.93 (m, 1H), 1.88-1.37 (m, 10H).

Example 43: 3-(5-(((1S,2S)-2-(diethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-59)

To a vial was added 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 100 mg, 0.27 mmol), acetaldehyde (2-1, 76 μL, 1.4 mmol), and DMF (2.7 mL). The resulting mixture was stirred for 15 min under an N₂ atmosphere and sodium triacetoxyborohydride (285 mg, 1.35 mmol) was then added. The reaction mixture was stirred at rt for 4 h, added to a saturated aqueous bicarbonate solution, and extracted with EtOAc (×3). The organic phase was dried over MgSO₄, filtered, and concentrated to dryness. The crude product was purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; eluting with Water/MeCN with 0.1% Formic Acid; 75 ml/min, 1.5 mL injection; Gradient: 10-30% MeCN over 3.5 min) to afford the formate salt of I-59 (11 mg, 0.023 mmol, 36% yield), as a white solid. MS [M+H]⁺=428.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.59 (dd, J=8.4, 2.8 Hz, 1H), 7.15 (d, J=2.2 Hz, 1H), 7.02 (dt, J=8.4, 2.8 Hz, 1H), 5.11-5.01 (m, 1H), 4.63-4.58 (m, 1H), 4.43-4.19 (m, 2H), 2.97-2.85 (m, 2H), 2.64-2.52 (m, 2H), 2.43-2.31 (m, 3H), 2.02-1.79 (m, 3H), 1.76-1.64 (m, 3H), 1.61-1.23 (m, 6H), 0.91 (t, J=7.0 Hz, 6H).

Example 44: 3-(5-(((1S,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-60)

Step 1. 3-Hydroxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (44-2)

DMAP (60 mg, 0.49 mmol) and TEA (2.4 mL, 17 mmol) were added to a solution of 3-methylpentane-1,3,5-triol (44-1, 580 mg, 4.32 mmol) in MeCN (20 mL) and the resulting solution was placed under an inert atmosphere and cooled to 0° C. A solution of TsCl (1813 mg, 9.51 mmol) in MeCN (5 mL) was then added and the reaction mixture was stirred at 0° C. for 1 h, warmed to rt, and stirred at rt for 4 h. The reaction mixture was filtered and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-60% EtOAc in heptane to afford 44-2 (1.40 g, 3.16 mmol, 73% yield) as a light brown oil. MS [M+H₂O]⁺=460.3. ¹H NMR (400 MHz, Chloroform-d) δ 7.80-7.75 (m, 4H), 7.38-7.32 (m, 4H), 4.16 (t, J=6.5 Hz, 4H), 2.46 (s, 6H), 1.88-1.75 (m, 4H), 1.17 (s, 3H).

Step 2. 3-Methoxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (44-3)

Trimethyloxonium tetrafluoroborate (334 mg, 2.26 mmol) was added to a solution of 3-hydroxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (44-2, 500 mg, 1.13 mmol) and Proton-sponge® (484 mg, 2.26 mmol) in DCM and the resulting mixture was stirred at rt overnight. The reaction mixture was then filtered and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-60% EtOAc in heptane to afford 44-3 (300 mg, 0.657 mmol, 58% yield) as a colorless oil. MS [M+H₂O]⁺=474.5. ¹H NMR (400 MHz, Chloroform-d) δ 7.80-7.75 (m, 4H), 7.37-7.32 (m, 4H), 4.11-4.00 (m, 4H), 3.02 (s, 3H), 2.45 (s, 6H), 1.87-1.75 (m, 4H), 1.08 (s, 3H).

Step 3. 3-(5-(((1S,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-60)

DIPEA (0.18 mL, 1.0 mmol) was added to a solution of 3-methoxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (44-3, 184 mg, 0.404 mmol) and 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 75 mg, 0.20 mmol) in MeCN (1.5 mL) and the resulting mixture was stirred at 120° C. under μW irradiation for 4 h. The reaction mixture was concentrated to dryness. The crude residue was taken up in DCM and washed with saturated NaHCO₃ (aq) and the phases were separated. The aqueous phase was extracted with DCM and the combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-80% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-60 (39 mg, 0.073 mmol, 36% yield) as an off-white solid. MS [M+H]⁺=484.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.59 (d, J=8.3 Hz, 1H), 7.21-7.15 (m, 1H), 7.04 (d, J=8.0 Hz, 1H), 5.06 (dd, J=13.2, 5.1 Hz, 1H), 4.67 (s, 1H), 4.37 (dd, J=17.1, 6.8 Hz, 1H), 4.25 (dd, J=17.1, 5.3 Hz, 1H), 3.00 (s, 3H), 2.97-2.84 (m, 1H), 2.80-2.71 (m, 1H), 2.62-2.55 (m, 2H), 2.48-2.31 (m, 4H), 2.02-1.87 (m, 2H), 1.84-1.60 (m, 4H), 1.60-1.39 (m, 5H), 1.39-1.20 (m, 4H), 0.98 (s, 3H).

Example 45: 3-(5-(((1S,2S)-2-(isobutylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-61)

To a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 100 mg, 0.269 mmol) in TFE (2 mL) was added isobutyraldehyde (17-1a, 0.03 mL, 0.3 mmol) and the resulting mixture was stirred at rt for 30 minutes. Sodium triacetoxyborohydride (171 mg, 0.808 mmol) was added. The reaction mixture was stirred at rt overnight and then a saturated aqueous solution of sodium hydrogen carbonate (20 mL) was added. The aqueous phase was extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and the organic solvent was concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford a yellow solid. The obtained solid was repurified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N) in DCM (with 0.1% NEt₃) to afford a yellow solid which was again repurified via silica gel chromatography eluting with 0-15% i-PrOH (with 1% Et₃N) in DCM (with 1% Et₃N) to afford I-61 (24 mg, 0.053 mmol, 20% yield) as a pale yellow solid. MS [M+H]⁺=428.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.14 (d, J=2.2 Hz, 1H), 7.06-6.99 (m, 1H), 5.06 (dd, J=13.2, 5.0 Hz, 1H), 4.44-4.19 (m, 3H), 2.90 (ddd, J=18.1, 13.6, 5.4 Hz, 1H), 2.83-2.77 (m, 1H), 2.64-2.55 (m, 1H), 2.44-2.27 (m, 3H), 2.02-1.94 (m, 1H), 1.87-1.38 (br multiplets, 11H), 0.83 (dd, J=6.6, 2.3 Hz, 6H).

Example 46: 3-(1-oxo-5-(((1S,2S)-2-(propylamino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-62)

To a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT 40-5, 100 mg, 0.269 mmol) in TFE (3 mL) was added propionaldehyde (46-1, 0.02 mL, 0.3 mmol). The resulting mixture was stirred at rt for 30 minutes and then sodium triacetoxyborohydride (171 mg, 0.808 mmol) was added. The reaction mixture was stirred at rt for 5 hours and then added to a saturated aqueous solution of sodium hydrogen carbonate (20 mL). The aqueous phase was extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-62 (49 mg, 0.11 mmol, 42% yield) as a pale yellow solid. MS [M+H]⁺=414.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.14 (d, J=2.0 Hz, 1H), 7.06-6.99 (m, 1H), 5.06 (dd, J=13.2, 5.0 Hz, 1H), 4.44-4.20 (m, 3H), 2.89 (ddd, J=27.1, 14.0, 7.1 Hz, 1H), 2.88-2.80 (m, 1H), 2.65-2.52 (m, 2H), 2.48-2.28 (m, 2H), 2.04-1.92 (m, 1H), 1.86-1.43 (m, 10H), 1.42-1.31 (m, 2H), 0.83 (t, J=7.4 Hz, 3H).

Example 47: 3-(5-(((1S,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-63)

In a μW vial, 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 101.8 mg, 0.274 mmol) and 3,3-bis(bromomethyl)oxetane (47-1, 85 mg, 0.35 mmol) were dissolved in MeCN (1 mL) and DIPEA (0.15 mL, 0.86 mmol). The resulting mixture was stirred in a μW reactor at 120° C. for 6 h, transferred to a reaction vial with MeCN, and then concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford a yellow solid. The obtained solid was then purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid; 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford the formate salt of I-63 (38 mg, 0.075 mmol, 27% yield), as a white solid. MS [M+H]⁺=454.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.15 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.11 (s, 1H), 7.05-6.96 (m, 1H), 5.14-5.01 (m, 1H), 4.53 (s, 4H), 4.47-4.14 (m, 3H), 3.35 (d, J=7.24 Hz, 2H), 3.18 (d, J=7.1 Hz, 2H), 2.91 (ddd, J=17.7, 13.4, 5.2 Hz, 1H), 2.71-2.56 (m, 1H), 2.42-2.29 (m, 1H), 2.04-1.92 (m, 1H), 1.87-1.31 (m, 11H).

Example 48: 3-(5-(((1S,2S)-2-((((1r,4S)-4-methoxycyclohexyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-64)

Step 1. ((1R,4R)-4-Methoxycyclohexyl)methanol (48-2)

To a solution of trans-4-methoxycyclohexane-1-carboxylic acid (48-1, 1.00 g, 6.32 mmol) in dry THF (10 mL), under an atmosphere of nitrogen and cooled using an ice bath, was added lithium aluminum hydride 1M in THF (9.5 mL, 9.5 mmol) dropwise. The resulting mixture was stirred using an ice bath for 2 h, then allowed to warm to room temperature and stirred for 16 h. A solution of saturated aqueous potassium sodium tartrate (Rochelle's Salt) (150 mL) was then added with stirring. The reaction mixture was extracted with DCM (×4) and the combined organic phases were passed through a phase separating column and concentrated to dryness to afford 48-2 (903 mg, 6.26 mmol, 99% yield) as a colorless oil. The product was carried onto the next step without purification. ¹H NMR (400 MHz, DMSO-d₆) δ 4.36 (s, 1H), 3.21 (s, 3H), 3.19 (d, J=6.3 Hz, 2H), 3.02 (tt, J=10.7, 4.1 Hz, 1H), 2.07-1.91 (m, 2H), 1.80-1.66 (m, 2H), 1.36-1.21 (m, 1H), 1.11-0.96 (m, 2H), 0.87 (tdd, J=13.2, 11.6, 3.1 Hz, 2H).

Step 2. (1R,4R)-4-Methoxycyclohexane-1-carbaldehyde (48-3)

((1R,4R)-4-methoxycyclohexyl)methanol (48-2, 100 mg, 0.693 mmol) was dissolved in DCM (3 mL) and cooled using an ice bath. DMP (412 mg, 0.971 mmol) was then added followed by H₂O (0.02 mL, 0.9 mmol) and the resulting mixture was stirred at rt for 4 hours. A 1:1 mixture of a saturated aqueous solution of sodium thiosulphate (10 mL) and a saturated aqueous solution of sodium hydrogen carbonate (10 mL) was added and the mixture was extracted with DCM (×3). The combined organic phases were passed through a phase separating column and concentrated to dryness to afford a white solid. The crude material was suspended in excess diethyl ether, sonicated and filtered. The obtained solid was washed with diethyl ether (×2). The filtrate was then collected and concentrated to dryness to afford 48-3 (104 mg, 0.585 mmol, 84% yield) as a colorless oil. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 9.57 (d, J=1.2 Hz, 1H), 3.23 (s, 3H), 3.12-2.99 (m, 1H), 2.31-2.16 (m, 1H), 2.05-1.81 (m, 4H), 1.33-1.09 (m, 4H).

Step 3. 3-(5-(((1S,2S)-2-((((1R,4S)-4-methoxycyclohexyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-64)

To a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 100 mg, 0.269 mmol) in TFE (2 mL) was added (1r,4r)-4-methoxycyclohexane-1-carbaldehyde (48-3, 42.1 mg, 0.296 mmol). The resulting mixture was stirred at rt for 30 minutes and then sodium triacetoxyborohydride (171 mg, 0.808 mmol) was added. The reaction mixture was stirred at rt overnight and then a saturated aqueous solution of sodium hydrogen carbonate (20 mL) was added. The mixture was extracted with 20% i-PrOH in DCM (×3) and the combined organic phases were passed through a phase separating column and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-64 (50 mg, 0.095 mmol, 36% yield), as a pale yellow solid. MS [M+H]⁺=498.4. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.13 (d, J=2.1 Hz, 1H), 7.02 (dt, J=8.4, 1.6 Hz, 1H), 5.11-5.02 (m, 1H), 4.43-4.21 (m, 3H), 3.20 (s, 3H), 3.05-2.84 (m, 2H), 2.83-2.74 (m, 1H), 2.64-2.54 (m, 1H), 2.45-2.27 (m, 3H), 2.02-1.90 (m, 3H), 1.85-1.58 (m, 7H), 1.58-1.38 (m, 5H), 1.33-1.21 (s, 1H), 1.02 (q, J=12.2 Hz, 2H), 0.93-0.76 (m, 2H).

Example 49: 3-(1-oxo-5-(((1S,2S)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cycloheptyl)oxy) isoindolin-2-yl)piperidine-2,6-dione (I-65)

To a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 100 mg, 0.269 mmol) in DCE (2 mL) was added tetrahydro-2H-pyran-4-carbaldehyde (49-1, 34 mg, 0.30 mmol). The resulting mixture was stirred at rt for 30 minutes, and then sodium triacetoxyborohydride (171 mg, 0.808 mmol) was added. The reaction mixture was stirred at rt overnight and then a saturated aqueous solution of sodium hydrogen carbonate (20 mL) was added. The aqueous mixture was extracted with 20% i-PrOH in DCM (×3) and the combined organics were passed through a phase separating column and concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid; 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-65 (43 mg, 0.075 mmol, 28% yield), as a white solid. MS [M+H]⁺=470.4. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.17-7.09 (m, 1H), 7.02 (dt, J=8.4, 1.8 Hz, 1H), 5.06 (dd, J=13.4, 4.9 Hz, 1H), 4.42-4.19 (m, 3H), 3.79 (dd, J=11.5, 4.0 Hz, 2H), 3.27-3.17 (m, 2H), 2.96-2.79 (m, 2H), 2.65-2.55 (m, 1H), 2.47-2.31 (m, 3H), 2.03-1.92 (m, 1H), 1.85-1.38 (m, 13H), 1.16-0.99 (m, 2H).

Example 50: 3-(5-(((1S,2S)-2-(((3-methyloxetan-3-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-66)

To a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 100 mg, 0.269 mmol) in DCE (3 mL) was added 3-methyloxetane-3-carbaldehyde (50-1, 32 mg, 0.32 mmol). The resulting mixture was stirred at rt for 30 minutes and then sodium triacetoxyborohydride (171 mg, 0.808 mmol) was added. The reaction mixture was stirred at rt overnight and then a saturated aqueous solution of sodium hydrogen carbonate (20 mL) was added. The aqueous mixture was extracted with 20% i-PrOH in DCM (×3) and the combined organics were passed through a phase separating column and concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid; 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-66 (18 mg, 0.034 mmol, 13% yield) as a white solid. MS [M+H]⁺=456.4. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.15 (s, 1H), 7.09-6.98 (m, 1H), 5.06 (dd, J=13.2, 5.0 Hz, 1H), 4.45-4.20 (m, 5H), 4.13 (dd, J=5.5, 2.9 Hz, 2H), 2.98-2.84 (m, 1H), 2.84-2.54 (m, 5H), 2.37 (dd, J=13.2, 4.5 Hz, 1H), 2.02-1.92 (m, 1H), 1.86-1.40 (m, 9H), 1.18 (s, 3H).

Example 51: 3-(5-(((1S,2S)-2-(((3-fluorobicyclo[0.1.1]pentan-1-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-67) Step 1. 3-fluorobicyclo[1.1.1]pentane-1-yl)methanol (51-2)

To a solution of 3-fluorobicyclo[1.1.1]pentane-1-carboxylic acid (51-1, 1000 mg, 7.69 mmol) in THF (3 mL) under a nitrogen atmosphere at 0° C. was added 1M BH₃ in THF (9.2 mL, 9.2 mmol) and the resulting mixture was stirred overnight at rt. The reaction mixture was then quenched with MeOH (2 mL) and concentrated to dryness to afford 51-2 (890 mg, 7.66 mmol, 100% yield) as a colorless oil which was carried onto the next step without purification. ¹H NMR (400 MHz, DCM-d₂) δ 3.84 (s, 2H), 2.02 (d, J=2.7 Hz, 6H).

Step 2. 3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (51-3)

To a solution of (3-fluorobicyclo[1.1.1]pentan-1-yl)methanol (51-2, 890 mg, 7.66 mmol) in DCM (10 mL) was added DMP (4.87 g, 11.5 mmol) and the resulting mixture was stirred at rt for 6 h. The reaction mixture was diluted with Et₂O and filtered. The filtrate was concentrated to dryness to afford 51-3, which was carried onto the next step without purification (assumed quantitative yield). ¹H NMR (400 MHz, DMSO-d₆) δ 9.29 (d, J=6.4 Hz, 1H), 1.91 (d, J=2.7 Hz, 6H).

Step 3. 3-(5-(((1S,2S)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (51-4)

To a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 102 mg, 0.275 mmol) in DCE (0.9 mL) was added pre-filtered 3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (51-3, 38 mg, 0.33 mmol). The resulting mixture was stirred at rt for 15 minutes and then sodium triacetoxyborohydride (87 mg, 0.41 mmol) was added. The reaction mixture was stirred at rt for 18 h and then quenched with saturated aqueous sodium bicarbonate and extracted with 20% i-PrOH in DCM (×3). The combined organic layers were passed through a phase separator and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane to afford I-67 (52.4 mg, 0.107 mmol, 39% yield) as a white solid. MS [M+H]⁺=470.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.62 (d, J=8.3 Hz, 1H), 7.15 (d, J=2.2 Hz, 1H), 7.03 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (ddd, J=13.2, 5.2, 1.8 Hz, 1H), 4.50-4.18 (m, 3H), 2.99-2.72 (m, 4H), 2.64-2.55 (m, 1H), 2.44-2.35 (m, 1H), 2.04-1.93 (m, 1H), 1.89 (d, J=2.6 Hz, 6H), 1.82-1.39 (m, 10H).

Example 52: 3-(5-(((1S,2S)-2-(3-methoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-68) Step 1. 2-methoxypropane-1,3-diol (52-2)

To a solution of dimethyl 2-methoxymalonate (52-1, 1.7 mL, 12 mmol) in dry THF (70 mL), under an atmosphere of nitrogen and cooled in an ice bath, was added lithium aluminum hydride 1M in THF (14.8 mL, 14.8 mmol) dropwise. The resulting mixture was stirred in an ice bath and then warmed to warm to rt and stirred at rt overnight. The reaction mixture was cooled using an ice bath and sodium sulfate decahydrate (4.77 g, 14.8 mmol) was added portion wise under a stream of nitrogen with vigorous stirring to provide a granular suspension. The suspension was filtered and the resulting solid was washed with THF (×3). The filtrate was concentrated to dryness to afford 52-2 (629 mg, 12.3 mmol, 48% yield) as a colorless oil. ¹H NMR (400 MHz, DMSO-d₆) δ 4.47 (t, J=5.6 Hz, 2H), 3.50-3.33 (m, 4H), 3.31 (s, 3H), 3.12 (quint, J=5.1 Hz, 1H).

Step 2. 2-methoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) (52-3)

DMAP (77 mg, 0.63 mmol) and TEA (1.05 mL, 7.54 mmol) were added to a solution of 2-methoxypropane-1,3-diol (52-2, 200 mg, 1.89 mmol) in MeCN (5 mL). TsCl (898 mg, 4.71 mmol) was added and the resulting mixture was stirred at rt overnight under an atmosphere of nitrogen. The reaction mixture was concentrated to dryness and the obtained residue treated with diethyl ether. The obtained suspension was filtered and the solid was washed with diethyl ether (×3). The filtrate was collected and concentrated to dryness to afford 52-3 (771 mg, 1.86 mmol, 99% yield) as a pale yellow oil. MS [M+H]⁺=415.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.78-7.74 (m, 4H), 7.35 (dd, J=8.7, 0.7 Hz, 4H), 4.07-3.97 (m, 4H), 3.59 (quint, J=5.0 Hz, 1H), 3.28 (s, 3H), 2.46 (s, 6H).

Step 3. 3-(5-(((1S,2S)-2-(3-methoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-68)

To a cloudy solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 100 mg, 0.269 mmol) in MeCN (2 mL) was added 2-methoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) (52-3, 167 mg, 0.404 mmol) and DIPEA (0.28 mL, 1.6 mmol). The resulting mixture was stirred at 120° C. for 2 hours and then at 140° C. for 1 hour in the μW. A saturated aqueous solution of sodium hydrogen carbonate (20 mL) and water (20 mL) was then added and the resulting mixture was extracted with 20% i-PrOH in DCM (×3). The combined organics were passed through a phase separating column and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-68 (40 mg, 0.09 mmol, 32% yield), as a pale yellow solid. MS [M+H]⁺=442.3. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.12 (s, 1H), 7.00 (dd, J=8.5, 2.2 Hz, 1H), 5.13-4.99 (m, 1H), 4.44-4.20 (m, 3H), 3.84 (t, J=5.8 Hz, 1H), 3.53-3.46 (m, 1H), 3.39-3.33 (m, 1H), 3.10 (s, 3H), 2.96-2.85 (m, 2H), 2.80-2.73 (m, 1H), 2.64-2.52 (m, 2H), 2.46-2.28 (m, 1H), 2.04-1.93 (m, 1H), 1.88-1.34 (m, 10H).

Example 53: 3-(5-(((1S,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-69)

Step 1. (3,3-difluorocyclobutyl)methyl 4-methylbenzenesulfonate (53-2)

To a solution of (3,3-difluorocyclobutyl)methanol (53-1, 100 mg, 0.82 mmol) in DCM (3 mL) was added DIPEA (0.29 mL, 1.6 mmol), 1-methyl-1H-imidazole (0.13 mL, 1.6 mmol), followed by TsCl (234 mg, 1.23 mmol) dropwise, under a stream of nitrogen. The resulting mixture was stirred at rt overnight and then diluted with DCM (30 mL total). The organic phase was separated and washed with 1M HCl (aq) (×2), saturated aqueous solution of sodium hydrogen carbonate (×2) and brine (×1). The organic phase was passed through a phase separating column and concentrated to dryness to afford crude 53-2 (236 mg) as a white solid. The crude product was carried onto the next step without purification. MS [M+H₂O]⁺=294.1. ¹H NMR (400 MHz, DMSO-d₆) δ 7.83-7.77 (m, 2H), 7.53-7.47 (m, 2H), 4.09 (dt, J=6.6, 0.9 Hz, 2H), 2.67-2.53 (m, 2H), 2.48-2.38 (m, 1H), 2.43 (s, 3H), 2.37-2.21 (m, 2H).

Step 2. 3-(5-(((1S,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-69)

To a suspension of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 112 mg, 0.302 mmol) in MeCN (2 mL), whilst bubbling with nitrogen, was added crude (3,3-difluorocyclobutyl)methyl 4-methylbenzenesulfonate (53-2, 92 mg, 0.33 mmol) and DIPEA (0.26 mL, 1.5 mmol). The resulting mixture was stirred at 120° C. for 2 hours and then at 140° C. for 9 hours in the μW. The reaction mixture was diluted with water (15 mL) and a saturated aqueous solution of sodium hydrogen carbonate (15 mL). The aqueous mixture was extracted with 20% i-PrOH in DCM (×3) and the combined organic phases were passed through a phase separating column and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-15% i-PrOH in DCM using 0.1% NEt₃ as a modifier, to afford I-69 (39 mg, 0.078 mmol, 26% yield) as a white solid. MS [M+H]⁺=476.3. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.14 (d, J=2.2 Hz, 1H), 7.03 (dd, J=8.4, 2.2 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.43-4.20 (m, 3H), 2.90 (ddd, J=18.1, 13.5, 5.3 Hz, 1H), 2.84-2.78 (m, 1H), 2.71-2.52 (m, 5H), 2.44-2.30 (m, 1H), 2.27-2.12 (m, 3H), 2.02-1.93 (m, 1H), 1.84-1.39 (m, 10H).

Example 54: 3-(5-(((1S,2S)-2-(3-ethoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-55)

Step 1. 2-Ethoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) (54-1)

TsCl (1368 mg, 7.18 mmol) was added to a solution of 2-ethoxypropane-1,3-diol (54-2, 392 mg, 3.26 mmol), DMAP (40 mg, 0.33 mmol), and TEA (1.8 mL, 13 mmol) in MeCN (10 mL). The resulting mixture was stirred at rt overnight and then filtered. The filtrate was collected and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc in heptane to afford 54-1 (220 mg, 0.513 mmol, 16% yield) as a colorless oil. MS [M+H₂O]⁺=446.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.78-7.72 (m, 4H), 7.38-7.32 (m, 4H), 4.05-3.96 (m, 4H), 3.68 (quint, J=5.1 Hz, 1H), 3.46 (q, J=7.0 Hz, 2H), 2.46 (s, 6H), 1.07 (t, J=7.0 Hz, 3H).

Step 3. 3-(5-(((1S,2S)-2-(3-ethoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-55)

DIPEA (0.17 mL, 0.97 mmol) was added to a solution of 3-(5-(((1S,2S)-2-aminocycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (INT-40-5, 60 mg, 0.16 mmol) and 2-ethoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) (54-1, 220 mg, 0.51 mmol) in MeCN (2 mL). The resulting mixture was stirred at 120° C. under μW irradiation for 3 h and then concentrated to dryness. The crude residue was triturated with diethyl ether (3×). The crude product was purified via silica gel chromatography eluting 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃). The obtained material was treated with 1:1 water/sat. aqueous Na₂CO₃ and extracted with DCM (×3). The combined organic phases were passed through a phase separating column and the solvent concentrated to dryness to afford I-55 (17 mg, 0.035 mmol, 22% yield) as an off-white solid. MS [M+H]⁺=456.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.12 (d, J=2.1 Hz, 1H), 7.00 (dd, J=8.7, 2.0 Hz, 1H), 5.07 (dd, J=13.2, 5.1 Hz, 1H), 4.39 (dd, J=17.1, 7.4 Hz, 2H), 4.25 (dd, J=17.2, 5.5 Hz, 1H), 3.91 (quint, J=5.9 Hz, 1H), 3.54-3.46 (m, 1H), 3.38-3.33 (m, 1H), 3.30-3.26 (m, 2H), 2.97-2.82 (m, 2H), 2.75 (t, J=6.4 Hz, 1H), 2.63-2.53 (m, 2H), 2.44-2.35 (m, 1H), 2.00-1.93 (m, 1H), 1.88-1.78 (m, 1H), 1.76-1.55 (m, 4H), 1.56-1.34 (m, 5H), 1.05 (t, J=7.0 Hz, 3H).

Example 55: 3-(1-oxo-5-(((1S,2S)-2-(propylamino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-71)

3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 121.8 mg, 0.34 mmol) and propionaldehyde (46-1, 0.04 mL, 0.6 mmol) were dissolved in 2,2,2-trifluoroethanol (1 mL). Sodium triacetoxyborohydride (108 mg, 0.511 mmol) was added in one portion and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was then diluted with ethyl acetate (45 mL) and washed with saturated aqueous sodium bicarbonate solution (10 mL) and brine (10 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collected fractions into tubes containing ˜3 drops of formic acid) to afford the formate salt of I-71 (36 mg, 0.078 mmol, 23% yield), as a solid. MS [M+H]⁺=400.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.20 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.21 (s, 1H), 7.08 (ddd, J=8.5, 2.3, 1.0 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.46-4.15 (m, 3H), 2.90 (ddd, J=17.2, 13.6, 5.4 Hz, 1H), 2.77-2.69 (m, 1H), 2.64-2.56 (m, 2H), 2.45-2.35 (m, 2H), 2.12-2.04 (m, 1H), 2.03-1.91 (m, 2H), 1.74-1.59 (m, 2H), 1.48-1.14 (m, 6H), 0.84 (t, J=7.4 Hz, 3H).

Example 56: 3-(5-(((1S,2S)-2-(dipropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-72)

3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 121.8 mg, 0.341 mmol) and propionaldehyde (46-1, 0.04 mL, 0.6 mmol) were dissolved in 2,2,2-trifluoroethanol (1 mL). Sodium triacetoxyborohydride (108 mg, 0.511 mmol) was added in one portion and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with ethyl acetate (45 mL) and washed with saturated aqueous sodium bicarbonate solution (10 mL) and brine (10 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 m 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collected fractions collected fractions into tubes containing ˜3 drops of formic acid) to afford crude material. Crude material was purified via silica gel chromatography eluting with 0-60% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N) to afford I-72 (26 mg, 0.060 mmol, 18% yield) as a solid. MS [M+H]⁺=442.6 m/z. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.14 (s, 1H), 6.99 (dd, J=8.5, 2.3, 1.1 Hz, 1H), 5.06 (ddd, J=13.2, 5.1, 2.4 Hz, 1H), 4.44 (q, J=5.0 Hz, 1H), 4.41-4.19 (m, 2H), 2.90 (ddd, J=17.3, 13.7, 5.4 Hz, 1H), 2.69-2.55 (m, 2H), 2.48-2.36 (m, 5H), 2.14 (d, J=11.0 Hz, 1H), 2.05-1.91 (m, 1H), 1.83-1.57 (m, 3H), 1.44-1.20 (m, 8H), 0.76 (t, J=7.3 Hz, 6H).

Example 57: 3-(1-oxo-5-(((1S,2S)-2-((pyridin-4-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-73)

3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 109.1 mg, 0.305 mmol) and isonicotinaldehyde (57-1, 0.03 mL, 0.3 mmol) were dissolved in 2,2,2-trifluoroethanol (1 mL), and sodium triacetoxyborohydride (83.2 mg, 0.393 mmol) was added in one portion. The resulting mixture was stirred at rt for 1 h. The reaction mixture was concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0-70% EtOAc:EtOH ((v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N). The obtained material was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-73 (21 mg, 0.042 mmol, 14% yield) as a white solid. MS [M+H]⁺=449.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.50-8.43 (m, 2H), 8.14 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.39-7.29 (m, 2H), 7.23-7.16 (m, 1H), 7.07 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (ddd, J=13.1, 5.0, 1.5 Hz, 1H), 4.47-4.19 (m, 3H), 3.91-3.75 (m, 2H), 2.91 (ddd, J=17.1, 13.5, 5.3 Hz, 1H), 2.67-2.55 (m, 2H), 2.44-2.34 (m, 1H), 2.07 (s, 1H), 2.03-1.89 (m, 2H), 1.64 (s, 2H), 1.45-1.13 (m, 4H).

Example 58: 3-(5-(((1S,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-74)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 90.3 mg, 0.253 mmol) dissolved in DMF (2 mL) was added DIPEA (0.11 mL, 0.63 mmol) followed by a solution of 3,3-bis(bromomethyl)oxetane (47-1, 73.6 mg, 0.302 mmol) in DMF (1 mL). The resulting mixture was stirred at 60° C. for 40 h. The reaction mixture was concentrated to dryness and the crude product purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N) to afford I-74 (10 mg, 0.022 mmol, 9% yield), as a pale yellow solid. MS [M+H]⁺=440.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.16 (s, 1H), 7.08-6.97 (m, 1H), 5.07 (dd, J=13.0, 5.0 Hz, 1H), 4.54 (t, J=4.9 Hz, 4H), 4.45-4.32 (m, 1H), 4.25 (dd, J=25.4, 8.3 Hz, 2H), 3.37 (d, J=7.5 Hz, 2H), 3.25 (d, J=7.6 Hz, 2H), 2.91 (ddd, J=18.5, 13.6, 5.3 Hz, 1H), 2.59 (d, J=16.7 Hz, 1H), 2.43-2.34 (m, 1H), 2.25 (s, 1H), 1.99 (s, 2H), 1.82-1.70 (m, 1H), 1.61 (d, J=9.9 Hz, 2H), 1.39-1.26 (m, 2H), 1.19 (s, 1H), 1.07-0.99 (m, 1H).

Example 59: 3-(1-oxo-5-(((1S,2S)-2-((pyridin-3-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-75)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 96.6 mg, 0.270 mmol) and 3-(bromomethyl)pyridine hydrobromide (59-1, 79.8 mg, 0.315 mmol) dissolved in MeCN (2 mL) was added DIPEA (0.15 mL, 0.87 mmol) and the resulting mixture was stirred at 60° C. for 4 days. The reaction mixture was concentrated to dryness and the crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collection tubes contain ˜3 drops of formic acid). The obtained material was further purified via silica gel chromatography eluting with 5-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N) to afford I-75 (17 mg, 0.039 mmol, 14% yield), as a white solid. MS [M+H]⁺=449.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.51 (s, 1H), 8.41 (d, J=4.7 Hz, 1H), 7.71 (d, J=7.8 Hz, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.31 (dd, J=7.7, 4.7 Hz, 1H), 7.19 (s, 1H), 7.07 (dd, J=8.4, 2.2 Hz, 1H), 5.13-5.01 (m, 1H), 4.44-4.18 (m, 3H), 3.82 (q, J=14.2 Hz, 2H), 2.90 (ddd, J=17.1, 13.6, 5.4 Hz, 1H), 2.70-2.55 (m, 2H), 2.46-2.29 (m, 1H), 2.10-2.03 (m, 1H), 2.03-1.91 (m, 2H), 1.74-1.57 (m, 2H), 1.46-1.17 (m, 4H).

Example 60: 3-(5-(((1S,2S)-2-(((1-ethyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-76)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 118.3 mg, 0.331 mmol) and 1-ethyl-1H-pyrazole-4-carbaldehyde (60-1, 46.1 mg, 0.371 mmol) in 2,2,2-trifluoroethanol (1.5 mL) was added sodium triacetoxyborohydride (98 mg, 0.46 mmol) in one portion. The resulting mixture was stirred at rt for 1 h and then diluted with ethyl acetate (80 mL) and washed with saturated aqueous sodium bicarbonate solution (10 mL) and brine (10 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated. The crude material was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford the formate salt of I-76 (54 mg, 0.10 mmol, 32% yield) as a white solid. MS [M+H]⁺=466.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.15 (s, 2H), 7.68-7.53 (m, 2H), 7.34 (s, 1H), 7.20 (s, 1H), 7.08 (dd, J=8.5, 2.1 Hz, 1H), 5.07 (dd, J=13.2, 5.1 Hz, 1H), 4.47-4.19 (m, 3H), 4.06 (q, J=7.2 Hz, 2H), 3.80-3.62 (m, 2H), 2.90 (ddd, J=18.2, 13.7, 5.5 Hz, 1H), 2.80 (s, 1H), 2.59 (d, J=17.4 Hz, 1H), 2.41-2.33 (m, 1H), 2.08 (s, 1H), 2.06-1.93 (m, 2H), 1.67 (s, 2H), 1.46-1.17 (m, 7H).

Example 61: 3-(5-(((1S,2S)-2-(((1-isopropyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-77)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 113.3 mg, 0.317 mmol) and 1-isopropyl-1H-pyrazole-4-carbaldehyde (61-1, 47.2 mg, 0.342 mmol) in 2,2,2-trifluoroethanol (1.5 mL) was added sodium triacetoxyborohydride (85.7 mg, 0.404 mmol) in one portion. The resulting mixture was stirred at rt for 2 h and then concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 5-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N) followed by purification via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford the formate salt of I-77 (57 mg, 0.104 mmol, 33% yield), as a white solid. MS [M+H]⁺=480.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.16 (s, 2H), 7.66-7.54 (m, 2H), 7.32 (d, J=0.9 Hz, 1H), 7.20 (d, J=2.2 Hz, 1H), 7.08 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.50-4.17 (m, 4H), 3.79-3.60 (m, 2H), 2.90 (ddd, J=17.2, 13.6, 5.4 Hz, 1H), 2.78 (d, J=5.8 Hz, 1H), 2.63-2.57 (m, 1H), 2.42-2.33 (m, 1H), 2.10-2.06 (m, 1H), 2.05-1.94 (m, 2H), 1.67 (s, 2H), 1.43-1.15 (m, 10H).

Example 62: 3-(5-(((1S,2S)-2-(ethyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-78)

Paraformaldehyde (6.23 mg, 0.207 mmol) and sodium triacetoxyborohydride (44.5 mg, 0.210 mmol) were added sequentially to a solution of 3-(5-(((1S,2S)-2-(ethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-79, 54 mg, 0.14 mmol) in DCM (1 mL) and the resulting mixture was stirred at rt. Acetic acid (2 μL, 0.03 mmol) was then added followed by an additional portion of sodium triacetoxyborohydride (44.5 mg, 0.210 mmol). The reaction mixture was stirred at rt for 16 h and then saturated NaHCO₃ (aq) (2 mL) was added. The mixture was extracted with DCM (×3) and the combined organic phases were concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-20% i-PrOH in DCM using 0.1% TEA as a modifier, to afford I-78 (38 mg, 0.090 mmol, 64% yield), as a white solid. MS [M+H]⁺=400.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.3 Hz, 1H), 7.17 (s, 1H), 7.03 (d, J=8.4 Hz, 1H), 5.06 (dd, J=13.4, 5.1 Hz, 1H), 4.55-4.44 (m, 1H), 4.38 (dd, J=17.0, 10.1 Hz, 1H), 4.25 (dd, J=17.0, 9.4 Hz, 1H), 2.97-2.85 (m, 1H), 2.74-2.64 (m, 1H), 2.63-2.52 (m, 3H), 2.44-2.34 (m, 1H), 2.19 (s, 3H), 2.15-2.06 (m, 1H), 2.03-1.93 (m, 1H), 1.79-1.58 (m, 3H), 1.43-1.20 (m, 4H), 0.91 (t, J=6.3 Hz, 3H).

Example 63: 3-(5-(((1S,2S)-2-(dimethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-80)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 75 mg, 0.18 mmol) in DCE (1 mL) was added paraformaldehyde (14 mg, 0.47 mmol) followed by sodium triacetoxyborohydride (111 mg, 0.524 mmol) and acetic acid (2 μL, 0.04 mmol) was added and the resulting mixture was stirred at rt for 16 h. The reaction mixture was poured into sat. NaHCO₃ (aq) (2 mL) and extracted with DCM (×4). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-20% i-PrOH in DCM using 0.1% Et₃N as a modifier. The obtained material was further purified via silica gel chromatography eluting with 0-10% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N) to afford I-80 (32 mg, 0.079 mmol, 43% yield) as a yellow solid. MS [M+H]⁺=386.5. ¹H NMR (400 MHz, DMSO-d₆) 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.19 (s, 1H), 7.11-6.99 (m, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.55-4.44 (m, 1H), 4.38 (dd, J=17.1, 8.8 Hz, 1H), 4.25 (dd, J=17.1, 8.2 Hz, 1H), 2.97-2.84 (m, 1H), 2.64-2.55 (m, 2H), 2.43-2.35 (m, 1H), 2.27 (s, 6H), 2.14-2.06 (m, 1H), 2.02-1.93 (m, 1H), 1.84-1.74 (m, 1H), 1.74-1.60 (m, 2H), 1.43-1.20 (m, 4H).

Example 64: 3-(5-(((1S,2S)-2-((oxetan-3-ylmethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-81)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 102.7 mg, 0.287 mmol) and 3-(bromomethyl)oxetane (64-1, 55 mg, 0.36 mmol) in DMF (1 mL) was added DIPEA (0.13 mL, 0.72 mmol) and the resulting mixture was stirred at 60° C. for 46 h. The reaction mixture was diluted with ethyl acetate (80 mL) and washed with saturated aqueous sodium bicarbonate solution (15 mL) and brine (15 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 5 to 100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N) and then further purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-81 (32 mg, 0.066 mmol, 23% yield) as a white solid. MS [M+H]⁺=428.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.14 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.20 (d, J=2.1 Hz, 1H), 7.08 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.57 (dd, J=7.4, 5.9 Hz, 2H), 4.38 (dd, J=17.2, 7.4 Hz, 1H), 4.31-4.14 (m, 4H), 3.04-2.82 (m, 4H), 2.77-2.66 (m, 1H), 2.59 (d, J=16.9 Hz, 1H), 2.38 (dd, J=13.2, 4.4 Hz, 1H), 2.10-2.03 (m, 1H), 2.03-1.92 (m, 2H), 1.71-1.62 (s, 2H), 1.40-1.17 (m, 4H).

Example 65: 3-(5-(((1S,2S)-2-((2-hydroxyethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-82)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 103 mg, 0.287 mmol) and 2-iodoethanol (65-1, 26 μL, 0.33 mmol) in DMF (1 mL) was added DIPEA (125 μL, 0.718 mmol) and the resulting mixture was stirred at 60° C. for 44 h. The reaction mixture was then diluted with ethyl acetate (80 mL) and washed with brine (2×15 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) and then further purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient: Collection tubes contain ˜3 drops of formic acid) to afford the formate salt of I-82 (18 mg, 0.038 mmol, 13% yield) as a white solid. MS [M+H]⁺=402.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.17 (s, 2H), 7.61 (d, J=8.4 Hz, 1H), 7.21 (s, 1H), 7.07 (dd, J=8.4, 2.3 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.38 (dd, J=17.2, 6.8 Hz, 1H), 4.26 (dd, J=17.2, 5.7 Hz, 2H), 3.47 (dt, J=10.5, 5.2 Hz, 2H), 2.90 (ddd, J=18.1, 13.6, 5.4 Hz, 1H), 2.76-2.67 (m, 2H), 2.65-2.57 (m, 1H), 2.38 (dd, J=13.2, 4.4 Hz, 1H), 2.07 (s, 1H), 2.03-1.92 (m, 2H), 1.66 (s, 2H), 1.26 (ddt, J=53.3, 21.2, 11.1 Hz, 4H).

Example 66: 3-(1-oxo-5-(((1S,2S)-2-(pyrrolidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-83)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 101 mg, 0.282 mmol) in DMF (2 mL) and DIPEA (0.12 mL, 0.69 mmol) was added a solution of 1,4-dibromobutane (66-1, 67.2 mg, 0.311 mmol) in DMF (1 mL) and the resulting mixture was stirred at 80° C. for 69 h. The reaction mixture was then diluted with ethyl acetate (80 mL) and washed with saturated sodium bicarbonate solution (10 mL) and brine (10 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-80% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM (with 1% Et₃N) and then further purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-83 (23 mg, 0.049 mmol, 18% yield) as a white solid. MS [M+H]⁺=412.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.15 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.20 (d, J=2.2 Hz, 1H), 7.07 (dd, J=8.3, 2.2 Hz, 1H), 5.06 (dd, J=13.3, 5.0 Hz, 1H), 4.60 (td, J=6.9, 3.4 Hz, 1H), 4.45-4.18 (m, 2H), 2.90 (ddd, J=18.5, 13.7, 5.4 Hz, 1H), 2.76-2.54 (m, 6H), 2.42-2.33 (m, 1H), 1.98 (td, J=7.2, 6.5, 2.7 Hz, 2H), 1.93-1.84 (m, 1H), 1.74-1.57 (m, 6H), 1.56-1.44 (m, 2H), 1.43-1.26 (m, 2H).

Example 67: 3-(5-(((1S,2S)-2-morpholinocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione I-84)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 75 mg, 0.183 mmol) in MeCN (1 mL) was added DIPEA (0.16 mL, 0.91 mmol) followed by 1-bromo-2-(2-bromoethoxy)ethane (29-1, 127 mg, 0.548 mmol) and the resulting mixture was stirred at 80° C. for 16 h. The crude reaction mixture was filtered, diluted with several drops of water, and purified by directly injecting it onto reverse phase preparative HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-84 (23 mg, 0.046 mmol, 25% yield) as a white solid. MS [M+H]⁺=428.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.20 (s, 1H), 7.06 (d, J=8.5 Hz, 1H), 5.07 (dd, J=13.4, 4.9 Hz, 1H), 4.55-4.46 (m, 1H), 4.39 (dd, J=17.0, 10.8 Hz, 1H), 4.26 (dd, J=17.1, 11.2 Hz, 1H), 3.46-3.34 (m, 4H), 2.97-2.84 (m, 1H), 2.70-2.54 (m, 4H), 2.43-2.35 (m, 3H), 2.16-2.07 (m, 1H), 2.03-1.93 (m, 1H), 1.82-1.74 (m, 1H), 1.72-1.58 (m, 2H), 1.46-1.20 (m, 4H).

Example 68: 3-(1-oxo-5-(((1S,2S)-2-((pyridin-2-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-85)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 63.5 mg, 0.18 mmol) and picolinaldehyde (68-1, 20 mg, 0.18 mmol) in 2,2,2-trifluoroethanol (1 mL) was added sodium triacetoxyborohydride (53 mg, 0.25 mmol) in one portion and the resulting mixture was stirred at rt for 1 h. The reaction solution was diluted with ethyl acetate (80 mL) and washed with saturated sodium bicarbonate solution (10 mL) and brine (10 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via reverse phase preparative HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) and then further purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-85 (35 mg, 0.070 mmol, 39% yield) as a solid. MS [M+H]⁺=449.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.48 (ddd, J=4.9, 1.8, 1.0 Hz, 1H), 8.14 (s, 1H), 7.72 (td, J=7.7, 1.8 Hz, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.40 (d, J=7.8 Hz, 1H), 7.26-7.17 (m, 2H), 7.08 (dd, J=8.6, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.45-4.19 (m, 3H), 4.01-3.79 (m, 2H), 2.91 (ddd, J=18.1, 13.6, 5.5 Hz, 1H), 2.73 (s, 1H), 2.59 (d, J=17.1 Hz, 1H), 2.40 (td, J=13.1, 4.4 Hz, 1H), 2.11-2.07 (m, 1H), 2.04-1.96 (m, 2H), 1.75-1.57 (m, 2H), 1.42-1.20 (m, 4H).

Example 69: 3-(5-(((1S,2S)-2-((3-hydroxy-3-methylbutyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-86)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 80 mg, 0.22 mmol) in MeCN (3 mL) was added 4-bromo-2-methylbutan-2-ol (69-1, 37.4 mg, 0.22 mmol) and DIPEA (0.12 mL, 0.67 mmol) and the resulting mixture was stirred at 65° C. for 16 h, at 100° C. for 30 minutes in the μW, at 120° C. for 30 minutes in the μW, and at 140° C. for 2 h and 30 minutes in the μW. The reaction mixture was then added to a saturated solution of sodium hydrogen carbonate (20 mL) and extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and then concentrated to dryness. The crude material was purified via reverse phase chromatography on C-18 column, eluting with 0-100% MeCN in water (with 0.1% Formic acid as a modifier). The fractions containing the desired product were combined and lyophilized to afford I-86 (16 mg, 0.031 mmol, 14% yield), as a white solid. MS [M+H]⁺=444.6. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm) 10.95 (s, 1H), 7.62 (d, J=8.5 Hz, 1H), 7.21 (s, 1H), 7.08 (d, J=8.5 Hz, 1H), 5.07 (dd, J=13.3, 5.2 Hz, 1H), 4.44-4.20 (m, 3H), 2.96-2.69 (m, 4H), 2.66-2.53 (m, 1H), 2.39 (td, J=13.2, 4.4 Hz, 1H), 2.10 (d, J=10.9 Hz, 1H), 2.06-1.94 (m, 2H), 1.72-1.64 (m, 2H), 1.52 (t, J=7.1 Hz, 2H), 1.42-1.19 (m, 4H), 1.07 (s, 6H).

Example 70: 3-(5-(((1S,2S)-2-(((3-methyloxetan-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-87)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 100 mg, 0.280 mmol) in DMF (1 mL) was added 3-methyloxetane-3-carbaldehyde (50-1, 34 mg, 0.34 mmol) and the resulting mixture was stirred at rt for 10 mins. Sodium triacetoxyborohydride (71 mg, 0.34 mmol) was then added and the reaction mixture was stirred at rt for 1 h, diluted with DCM, and Celite was added. The resulting suspension concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptane followed by 0 to 100% EtOAc/EtOH (v/v=3:1, with 1% NEt₃) in DCM and further purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 m 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collection tubes contained ˜3 drops of formic acid) to afford the formate salt of I-87 (40 mg, 0.080 mmol, 29% yield) as a white solid. MS [M+H]⁺=442.3. ¹H NMR (400 MHz, DMSO-d₆) δ 8.24 (s, 2H), 7.60 (d, J=8.4 Hz, 1H), 7.20 (s, 1H), 7.07 (dd, J=8.4, 2.1 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.37 (dd, J=17.1, 8.6 Hz, 1H), 4.30-4.17 (m, 4H), 4.11 (dd, J=5.4, 2.7 Hz, 2H), 2.96-2.85 (m, 1H), 2.80-2.68 (m, 2H), 2.68-2.53 (m, 2H), 2.35 (dd, J=15.9, 11.3 Hz, 1H), 2.11-2.04 (m, 1H), 2.02-1.92 (m, 2H), 1.71-1.62 (m, 2H), 1.41-1.19 (m, 4H), 1.15 (s, 3H).

Example 71: 3-(5-(((1S,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-88)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 29.7 mg, 0.083 mmol) in MeCN (1 mL) was added DIPEA (0.09 mL, 0.5 mmol) followed by 3-methoxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (44-3, 38 mg, 0.083 mmol) and the resulting mixture was stirred at 120° C. under μW irradiation for 2 h, then at 130° C. for 1 h in the μW. The reaction mixture was concentrated to dryness and the crude material was dissolved in DCM and washed with saturated NaHCO₃ (aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-88 (15 mg, 0.029 mmol, 35% yield) as a white solid. MS [M+H]⁺=470.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.59 (d, J=8.4 Hz, 1H), 7.18 (s, 1H), 7.04 (d, J=8.4 Hz, 1H), 5.07 (dd, J=13.3, 5.2 Hz, 1H), 4.52-4.42 (m, 1H), 4.37 (dd, J=17.2, 3.7 Hz, 1H), 4.25 (dd, J=17.1, 3.8 Hz, 1H), 3.33-3.31 (m, 2H, shoulder on H₂O signal), 2.97-2.93 (m, 3H), 2.93-2.85 (m, 1H), 2.63-2.55 (m, 3H), 2.45-2.35 (m, 2H), 2.15-2.08 (m, 1H), 2.03-1.94 (m, 1H), 1.79-1.58 (m, 3H), 1.53-1.42 (m, 2H), 1.40-1.04 (m, 6H), 0.92 (s, 3H)

Example 72: 3-(5-(((1S,2S)-2-(3-methoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-89)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 50 mg, 0.14 mmol) in MeCN (1.5 mL) was added DIPEA (0.15 mL, 0.84 mmol), followed by 2-methoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) (52-3, 174 mg, 0.42 mmol) and the resulting mixture was stirred at 120° C. under μW irradiation for 4 h. The reaction mixture was concentrated to dryness, the obtained residue dissolved in DCM and washed with saturated NaHCO₃(aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic layers were dried over Na₂SO₄, filtered and concentrated. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-89 (32 mg, 0.067 mmol, 48% yield) as a white solid. MS [M+H]⁺=428.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.16 (d, J=2.0 Hz, 1H), 7.02 (dd, J=8.4, 2.2 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.43-4.33 (m, 1H), 4.30-4.19 (m, 2H), 3.90-3.76 (m, 1H), 3.50-3.40 (m, 2H), 3.11 (s, 3H), 3.02-2.79 (m, 3H), 2.63-2.55 (m, 1H), 2.43-2.27 (m, 2H), 1.98 (d, J=10.3 Hz, 2H), 1.83-1.73 (m, 1H), 1.69-1.58 (m, 2H), 1.35 (t, J=8.8 Hz, 2H), 1.31-1.21 (m, 1H), 1.13-1.03 (m, 1H).

Example 73: 3-(5-(((1S,2S)-2-(((6-methylpyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-90)

Step 1. 2-(bromomethyl)-6-methylpyridine (73-2)

To 2,6-lutidine (73-1, 23 mL, 0.20 mol) in carbon tetrachloride (250 mL) was added NBS (35.6 g, 0.20 mol) and AIBN (0.5 g) to give a yellow suspension. The resulting suspension was stirred at reflux for 8 h to give a solution and then cooled to rt. The resultant suspension was filtered and the obtained solid was rinsed with carbon tetrachloride. The filtrate was collected and concentrated to dryness to afford 73-2 (33.7 g, 0.18 mol, 91% yield) as a violet colored oil which was carried onto the next step without purification. MS [M+H]⁺=185.6.

Step 2. 3-(5-(((1S,2S)-2-(((6-methylpyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-90)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 88.7 mg, 0.25 mmol) and 2-(bromomethyl)-6-methylpyridine (73-2, 50 mg, 0.269 mmol) in DMF (1 mL) was added DIPEA (0.12 mL, 0.69 mmol). The reaction mixture was stirred at 85° C. for 3 days. The reaction mixture was concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃). The obtained material was further purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford the formate salt of I-90 (4.0 mg, 7.47 μmol, 3% yield) as a yellow solid. MS [M+H]⁺=463.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.17 (s, 2H), 7.65-7.55 (m, 2H), 7.23-7.16 (m, 2H), 7.11-7.04 (m, 2H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.47-4.18 (m, 3H), 3.90-3.74 (m, 2H), 2.90 (ddd, J=17.8, 13.5, 5.4 Hz, 1H), 2.72-2.64 (m, 1H), 2.59 (d, J=17.1 Hz, 1H), 2.45-2.34 (m, 4H), 2.11-2.04 (m, 1H), 2.03-1.91 (m, 2H), 1.72-1.61 (m, 2H), 1.45-1.15 (m, 4H).

Example 74: 3-(5-(((1S,2S)-2-(((5-methoxypyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-91)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 106 mg, 0.30 mmol) and 2-formyl-5-methoxypyridine (74-1, 43 mg, 0.32 mmol) in 2,2,2-trifluoroethanol (1.5 mL) was added sodium triacetoxyborohydride (88 mg, 0.42 mmol) in one portion and the resulting mixture was stirred at room temperature for 90 min and then concentrated to dryness. The crude material was purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) and then further purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-91 (54.5 mg, 0.112 mmol, 38% yield) as a white solid. MS [M+H]⁺=479.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.19 (t, J=1.8 Hz, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.33 (d, J=1.8 Hz, 2H), 7.19 (s, 1H), 7.07 (dt, J=8.4, 2.0 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.48-4.16 (m, 3H), 3.92-3.83 (m, 1H), 3.83-3.75 (m, 4H), 2.91 (ddd, J=17.2, 13.6, 5.4 Hz, 1H), 2.79-2.65 (m, 1H), 2.59 (d, J=17.4 Hz, 1H), 2.45-2.28 (m, 1H), 2.11-1.95 (m, 3H), 1.74-1.59 (m, 2H), 1.45-1.21 (m, 4H).

Example 75: 3-(5-(((1S,2S)-2-(((6-methoxypyridin-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-92)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 75 mg, 0.210 mmol) and 6-methoxy-3-pyridinecarboxaldehyde (75-1, 30.5 mg, 0.222 mmol) in 2,2,2-trifluoroethanol (1 mL) was added sodium triacetoxyborohydride (58 mg, 0.27 mmol) in one portion and the resulting mixture was stirred at rt for 2 h and then concentrated to dryness. The crude material was purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 m 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford a formate salt of I-92 (34 mg, 0.063 mmol, 30% yield) as a white solid. MS [M+H]⁺=479.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.14 (s, 1H), 8.10-8.04 (m, 1H), 7.68-7.58 (m, 2H), 7.18 (s, 1H), 7.06 (dd, J=8.4, 2.2 Hz, 1H), 6.76 (dd, J=8.4, 0.7 Hz, 1H), 5.07 (dd, J=13.1, 5.1 Hz, 1H), 4.49-4.17 (m, 3H), 3.83-3.69 (m, 2H), 3.81 (s, 3H), 2.91 (ddd, J=18.2, 13.7, 5.5 Hz, 1H), 2.64-2.55 (m, 1H), 2.46-2.28 (m, 2H), 2.09-2.03 (m, 1H), 1.98 (t, J=5.1 Hz, 2H), 1.65 (d, J=9.9 Hz, 2H), 1.29 (dt, J=37.1, 9.2 Hz, 4H).

Example 76: 3-(5-(((1S,2S)-2-((2-hydroxy-2-methylpropyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-93)

2,2-dimethyloxirane (76-1, 0.015 mL, 0.17 mmol) and DIPEA (0.04 mL, 0.2 mmol) were added to a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 50 mg, 0.14 mmol) in EtOH (1.5 mL) and the resulting solution was stirred at 140° C. under μW irradiation for 1 h. Additional 2,2-dimethyloxirane (50 μL) was added and stirring was continued in the μW at 140° C. for 21 h. The reaction mixture was filtered and concentrated. The crude material was purified via reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collection tubes contained several drops of formic acid) to afford the formate salt of I-93 (12 mg, 0.023 mmol, 17% yield) as a white solid. MS [M+H]⁺=430.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.94 (br s, 1H), 8.22 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.21 (s, 1H), 7.06 (dd, J=8.4, 2.2 Hz, 1H), 5.06 (dd, J=13.3, 5.0 Hz, 1H), 4.38 (dd, J=17.2, 8.7 Hz, 1H), 4.30-4.19 (m, 3H), 2.96-2.84 (m, 1H), 2.66-2.54 (m, 1H), 2.47-2.35 (m, 3H), 2.12-2.03 (m, 1H), 2.03-1.91 (m, 2H), 1.73-1.60 (m, 2H), 1.42-1.12 (m, 4H), 1.03 (d, J=5.4 Hz, 6H).

Example 77: (1S,3r)-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile and (1R,3s)-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile (I-94 & I-95)

Step 1. 3-(Methoxymethylene)-1-methylcyclobutane-1-carbonitrile (77-3)

To a stirred suspension of (methoxymethyl)triphenylphosphonium chloride (77-2, 1828 mg, 5.33 mmol) in THF (30 mL) at 0° C. under an atmosphere of nitrogen was added n-BuLi (2.3 mL, 6.1 mmol, 2.7 M solution in heptane) and the resulting deep-red solution was stirred at 0° C. for 40 minutes. A solution of 1-methyl-3-oxocyclobutane-1-carbonitrile (77-1, 582 mg, 5.33 mmol) in THF (4 mL) was then added and the reaction mixture was stirred overnight, allowing the reaction to slowly warm to rt. The mixture was concentrated to dryness and the crude material purified via silica gel chromatography eluting with 0-100% EtOAc in diethyl ether to afford 77-3 (459 mg, 3.35 mmol, 63% yield) as a pale orange brown colored oil. ¹H NMR (400 MHz, DMSO-d₆) δ 6.10-5.94 (m, 1H), 3.51 (s, 3H), 3.19-2.97 (m, 2H), 2.81-2.57 (m, 2H), 1.47 (s, 3H).

Step 2. 3-Formyl-1-methylcyclobutane-1-carbonitrile (77-4)

To 3-(methoxymethylene)-1-methylcyclobutane-1-carbonitrile (77-3, 118 mg, 0.860 mmol) in DCM (2 mL) at rt was added trifluoroacetic acid (0.70 mL, 9.1 mmol) and water (100 μL, 5.6 mmol) and the resulting mixture was stirred at rt for 40 min and then saturated sodium bicarbonate. The aqueous mixture was extracted with DCM, the phases separated and the organic phase passed through a phase separating column. The organic solvent was evaporated to dryness to afford 77-4 as an oil, which was carried onto the next step without purification. ¹H NMR (400 MHz, DMSO-d₆) δ 9.66 (dd, J=10.5, 1.3 Hz, 1H), 3.45-3.30 (m, 1H), 2.75-2.56 (m, 2H), 2.39-2.11 (m, 2H), 1.53 (s, 3H).

Step 3. 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile (I-94)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 300 mg, 0.336 mmol) in TFE (1 mL) was added a solution of 3-formyl-1-methylcyclobutane-1-carbonitrile (77-4, 108 mg, 0.873 mmol) in TFE (1 mL). The resulting mixture was stirred at rt for 5 minutes and then sodium triacetoxyborohydride (213 mg, 1.00 mmol) was added. The reaction mixture was stirred at rt for 3 days and then concentrated to dryness. The crude material purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N as a modifier) in DCM to afford a mixture of I-94 and I-95 (40 mg, 0.082 mmol) as 66:34 mixture of diastereoisomers. MS [M+H]⁺=465.2.

The diastereomeric mixture of I-94 and I-95 (30 mg) was separated using chiral SFC (Column: Chiralpak IH 21×250 mm; Flow rate: 80 g per minute; Cosolvent: 30% IPA with 10 mM NH₃) to isolate the two diastereomers. Peak 1: diastereomer 1 (I-94, 13 mg, 0.026 mmol, 8% yield). Chiral SFC Rt=2.39 mins. MS [M+H]⁺=465.5. ¹H NMR (400 MHz, DCM-d₂) δ 7.71 (d, J=8.9 Hz, 1H), 7.11-6.89 (m, 2H), 5.11 (dd, J=13.4, 5.1 Hz, 1H), 4.40-4.23 (m, 2H), 4.21-4.05 (m, 1H), 2.85-2.77 (m, 2H), 2.74-2.47 (m, 4H), 2.32 (qd, J=12.8, 6.0 Hz, 1H), 2.25-2.17 (m, 2H), 2.17-2.04 (m, 4H), 1.79-1.71 (m, 2H), 1.49 (s, 2H), 1.40 (s, 1H), 1.38-1.22 (m, 5H), 1.20 (s, 1H). Peak 2: diastereomer 2 (I-95, 14 mg, 0.028 mmol, 8% yield). Chiral SFC Rt=2.70 mins. MS [M+H]⁺=465.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.70 (d, J=8.3 Hz, 1H), 7.17-6.92 (m, 2H), 5.11 (dd, J=13.5, 5.2 Hz, 1H), 4.34-4.24 (m, 2H), 4.24-4.16 (m, 1H), 2.88-2.65 (m, 5H), 2.62-2.52 (m, 1H), 2.41-2.03 (m, 7H), 1.80-1.71 (m, 2H), 1.49 (s, 2H), 1.40 (s, 1H), 1.37-1.24 (m, 5H), 1.21 (s, 1H). The relative stereochemistry of the cyclobutane substituents was not determined and was arbitrarily assigned.

Example 78: 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-96)

Step 1. 1,5-dichloropentan-3-ol (78-2)

To a solution of 1,5-dichloropentan-3-one (78-1, 260 mg, 1.68 mmol) in MeOH (2 mL) and cooled to −10° C. was added sodium borohydride (64 mg, 1.7 mmol) and the resulting mixture was stirred in an ice bath for 4 h. The reaction mixture was then poured into ice water and extracted with DCM (×3). The combined organic phases were washed with brine, passed through a phase separator, and concentrated to dryness to afford 78-2 (267 mg, 1.55 mmol, 92% yield) as a pale yellow oil. The product was carried onto the next step without purification. ¹H NMR (400 MHz, Chloroform-d) δ 4.14 (quint, J=6.2 Hz, 1H), 3.77-3.63 (m, 4H), 1.97-1.87 (m, 4H).

Step 2. 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-96)

To 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 52 mg, 0.11 mmol) dissolved in DMF (1 mL) was added DIPEA (0.10 mL, 0.57 mmol), followed by a solution of 1,5-dichloropentan-3-ol (78-2, 19 mg, 0.11 mmol) in DMF (1 mL) and the resulting mixture was stirred at 85° C. overnight. The reaction mixture was cooled to rt and additional 1,5-dichloropentan-3-ol (78-2, 19 mg, 0.11 mmol) was added and stirring was continued at 85° C. overnight. The reaction mixture was cooled to rt, azeotroped with toluene (×2), and concentrated. The crude material was purified by reverse phase HPLC (Method Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; fractions collected into tubes containing several drops of formic acid) to afford the formate salt of I-96 (9 mg, 0.018 mmol) as a pale brown solid. MS [M+H]⁺=428.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.15 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.1 Hz, 1H), 7.07-6.98 (m, 1H), 5.07 (dd, J=13.0, 4.9 Hz, 1H), 4.73-4.67 (m, 1H), 4.53-4.47 (m, 1H), 4.38 (dd, J=17.2, 9.8 Hz, 1H), 4.25 (dd, J=17.2, 8.3 Hz, 1H), 3.46-3.38 (m, 1H), 2.96-2.81 (m, 2H), 2.81-2.69 (m, 2H), 2.64-2.56 (m, 1H), 2.40-2.34 (m, 1H), 2.17-2.10 (m, 2H), 2.04-1.84 (m, 3H), 1.75-1.56 (m, 5H), 1.53-1.42 (m, 1H), 1.41-1.29 (m, 2H).

Example 79: 3-(1-oxo-5-(((1S,2S)-2-(piperidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-97)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 80 mg, 0.22 mmol) in DMF (3 mL) was added DIPEA (0.098 mL, 0.56 mmol) and 1,5-dibromopentane (79-1, 0.03 mL, 0.2 mmol) and the resulting mixture was stirred at 65° C. overnight. The reaction mixture was then cooled to rt and a saturated aqueous solution of sodium hydrogen carbonate (20 mL) was added. The aqueous mixture was extracted with 20% i-PrOH in DCM (×3). The combined organic phases were washed with water and brine, passed through a phase separating column, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% 3:1 EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-97 (25 mg, 0.056 mmol, 25% yield) as a white solid. MS [M+H]⁺=426.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (s, 1H), 7.04 (d, J=8.3 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.47 (br s, 1H), 4.42-4.18 (m, 2H), 2.90 (ddd, J=17.8, 13.2, 5.3 Hz, 1H), 2.69-2.44 (m, 6H), 2.39 (td, J=13.2, 4.5 Hz, 1H), 2.11 (s, 1H), 2.03-1.91 (m, 1H), 1.81-1.56 (m, 3H), 1.47-1.12 (m, 10H).

Example 80: 3-(1-oxo-5-(((1S,2S)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-98)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.15 g, 0.17 mmol) in DMF (1 mL), was added tetrahydro-2H-pyran-4-carbaldehyde (49-1, 0.021 mL, 0.21 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (55 mg, 0.26 mmol) was added and stirring was continued at rt for 16 h. Additional tetrahydro-2H-pyran-4-carbaldehyde (49-1, 0.01 mL, 0.1 mmol) was added and the resulting mixture was stirred at rt for 15 minutes. Additional sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added with stirring at rt for 1.5 h. The reaction mixture was diluted with dichloromethane and concentrated onto Celite®. The obtained material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane to afford I-98 (72.8 mg, 0.160 mmol, 93% yield) as a white solid. LCMS [M+H]⁺=456.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.20 (t, J=2.5 Hz, 1H), 7.06 (dt, J=8.5, 2.0 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.38 (dd, J=17.2, 8.2 Hz, 1H), 4.32-4.15 (m, 2H), 3.86-3.68 (m, 2H), 3.29-3.18 (m, 2H), 2.91 (ddd, J=17.5, 13.6, 5.4 Hz, 1H), 2.69-2.55 (m, 2H), 2.48-2.31 (m, 3H), 2.12-2.02 (m, 1H), 2.02-1.92 (m, 2H), 1.69-1.48 (m, 5H), 1.38-0.99 (m, 6H).

Example 81: 3-(5-(((1S,2S)-2-((((1R,3S)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1S,2S)-2-((((1R,3R)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-99 & I-100)

Step 1. (3-Methoxycyclobutyl)methanol (81-2)

To a solution of 3-methoxycyclobutane-1-carboxylic acid (81-1, 444 mg, 3.41 mmol) in THF (10 mL) at 0° C. was added BH₃ in THF (10.2 mL, 10.2 mmol, 1M) and the resulting mixture was allowed to warm slowly to rt and then stirred at rt for 18 h. MeOH was slowly added, followed by the addition of aqueous saturated bicarbonate solution. The aqueous mixture was extracted with DCM and the organic phase was passed through a phase collector and concentrated to dryness to afford 81-2. The crude material was taken through to the next step without purification.

Step 2. (3-Methoxycyclobutyl)methyl 4-methylbenzenesulfonate (81-3)

To a solution of (3-methoxycyclobutyl)methanol (81-2, 411 mg, 3.54 mmol) in pyridine (8 mL) was added TsCl (1012 mg, 5.31 mmol). The resulting mixture was stirred at rt for 2 h and then diluted with DCM and washed with a saturated aqueous bicarbonate solution. The organic phase was passed through a phase separating column and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc in heptanes to afford 81-3 (300 mg, 1.11 mmol, 31% yield), as a 2:1 mixture of diastereomers. ¹H NMR (400 MHz, DMSO-d₆, major diastereoisomer) δ 7.81-7.76 (m, 2H), 7.51-7.47 (m, 2H), 3.97 (d, J=6.1 Hz, 2H), 3.71-3.57 (m, 1H), 3.06 (s, 3H), 2.50 (quint, J=1.8 Hz, 1H), 2.43 (s, 3H), 2.22-2.15 (m, 2H), 1.48 (m, 2H). ¹H NMR (400 MHz, DMSO-d₆, minor diastereoisomer) δ 7.81-7.76 (m, 2H), 7.51-7.47 (m, 2H), 4.06-4.00 (m, 2H), 3.87-3.79 (m, 1H), 3.06 (s, 3H), 2.43 (s, 3H), 2.13-2.04 (m, 2H), 1.93-1.89 (m, 3H).

Step 3. 3-(5-(((1S,2S)-2-((((1R,3S)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1S,2S)-2-((((1R,3R)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-99) & (I-100)

To a vial containing 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 322 mg, 0.369 mmol), was added a solution of (3-methoxycyclobutyl)methyl 4-methylbenzenesulfonate (81-3, 300 mg, 1.11 mmol) in DMF (3 mL), followed by the addition of DIPEA (0.32 mL, 1.85 mmol). The reaction mixture was stirred at 45° C. for 16 h, then at 80° C. for 16 h. The reaction mixture was diluted with acetone, absorbed onto Celite® and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc in heptanes and then 0-100% EtOAc/EtOH (v/v=3:1) in DCM. The obtained material was purified further via reverse phase HPLC (10-30% MeCN, 3.5 min gradient X-bridge C18 OBD 30×50 mm 5 um column MeCN/H₂O w/0.1% Formic Acid 75 ml/min 1.5 mL injection) to afford the formate salt of a mixture of diastereomers I-99 and I-100 (87 mg, 0.17 mmol, 3% yield) as an oil. MS [M+H]⁺=456.5

The mixture of I-99 and I-100 (81 mg) was separated using chiral SFC (Column: Chiralpak AD 21×250 mm; Flow rate: 80 g per minute; Cosolvent: 25% EtOH with 10 mM NH₃) to afford I-99 and I-100 as Peak 1 and Peak 2. Peak 1 (I-99): The material from SFC Peak 1 was repurified via reverse phase HPLC (Method: eluting with 5-20% MeCN over 3.5 min; Conditions: MeCN/H₂O+0.1% Formic Acid at 75 mL/min; 1.5 mL injection; Column: XBridge C18 OBD 30×50 mm) and further purified via reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collection tubes contained several drops of formic acid) to afford the formate salt of Peak 1 (I-99, 9 mg, 0.028 mmol). Chiral SFC Rt=1.67 mins. MS [M+H]⁺=456.5. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 7.70 (d, J=9.1 Hz, 1H), 7.06-6.95 (m, 2H), 5.12 (dd, J=13.3, 5.2 Hz, 1H), 4.43-4.25 (m, 2H), 4.21-4.09 (m, 1H), 3.87 (quint, J=6.6 Hz, 1H), 3.14 (s, 3H), 2.88-2.79 (m, 2H), 2.76-2.68 (m, 2H), 2.68-2.59 (m, 1H), 2.38-2.24 (m, 2H), 2.23-2.03 (m, 5H), 2.01-1.87 (m, 5H), 1.77-1.70 (m, 2H), 1.37-1.33 (m, 2H). The relative stereochemistry of the cyclobutane substituents was not determined and was assigned arbitrarily.

Peak 2 (I-100): The material from SFC Peak 2 was purified via reverse phase HPLC (Method: eluting with 5-20% MeCN over 3.5 min. Conditions: MeCN/H₂O+0.1% Formic Acid at 75 mL/min; 1.5 mL injection; Column: XBridge C18 OBD 30×50 mm) to afford the formate salt of Peak 2 (I-100, 14 mg, 0.027 mmol). Chiral SFC Rt=2.52 mins. MS [M+H]⁺=456.6. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 7.70 (d, J=9.0 Hz, 1H), 7.08-6.98 (m, 2H), 5.23-5.01 (m, 1H), 4.36-4.24 (m, 2H), 4.15 (s, 1H), 3.76-3.62 (m, 1H), 3.15 (s, 3H), 2.86-2.69 (m, 3H), 2.64-2.60 (m, 1H), 2.36-2.26 (m, 2H), 2.22-2.10 (m, 3H), 2.09-2.02 (m, 1H), 2.00-1.85 (m, 2H), 1.79-1.68 (m, 2H), 1.54-1.43 (m, 1H), 1.38-1.31 (m, 2H), 1.31-1.22 (m, 3H). The relative stereochemistry of the cyclobutane substituents was not determined and was assigned arbitrarily.

Example 83: 3-(5-(((1S,2S)-2-((((1r,4S)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-101)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.16 g, 0.18 mmol) in DMF (1.1 mL) was added (1r,4r)-4-methoxycyclohexane-1-carbaldehyde (48-3, 37 mg, 0.26 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (58 mg, 0.28 mmol) was added and stirring was continued at rt for 1 h. Additional (1r,4r)-4-methoxycyclohexane-1-carbaldehyde (48-3, 7.3 mg, 0.051) was added and the resulting mixture was stirred at rt for 15 minutes. Additional sodium triacetoxyborohydride (25 mg, 0.12 mmol) was added and stirring was continued at rt for 16 h. The reaction mixture was quenched with sat. aqueous sodium bicarbonate and extracted with 20% isopropanol in DCM (×3). The organic phases were combined, passed through a phase separator, and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N as a modifier) in heptane and then purified further by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-101 (57 mg, 0.10 mmol, 55% yield) as a white solid. MS [M+H]⁺=484.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.19 (d, J=2.3 Hz, 1H), 7.06 (dt, J=8.4, 1.9 Hz, 1H), 5.07 (dd, J=13.3, 5.0 Hz, 1H), 4.38 (dd, J=17.3, 7.9 Hz, 1H), 4.31-4.14 (m, 2H), 3.19 (s, 3H), 3.05-2.84 (m, 2H), 2.70-2.55 (m, 2H), 2.46-2.29 (m, 3H), 2.13-1.88 (m, 5H), 1.82-1.59 (m, 4H), 1.40-1.10 (m, 5H), 1.07-0.95 (m, 2H), 0.91-0.75 (m, 2H).

Example 84: 3-(5-(((1S,2S)-2-(((4-methyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-102)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.16 g, 0.45 mmol) in DMF (0.92 mL) was added 4-methyltetrahydro-2H-pyran-4-carbaldehyde (84-1, 28 mg, 0.22 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (58 mg, 0.28 mmol) was added and stirring was continued at rt for 1 h. Additional 4-methyltetrahydro-2H-pyran-4-carbaldehyde (4.7 mg, 0.037 mmol) was added and the reaction mixture stirred at rt for 15 minutes. Additional sodium triacetoxyborohydride (19 mg, 0.092 mmol) was added and stirring was continued at rt for 16 h. The reaction mixture was then diluted with MeCN and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N as a modifier) in heptane to afford I-102 (99 mg, 0.21 mmol, 47% yield) as a white solid. MS [M+H]⁺=470.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.21 (d, J=2.0 Hz, 1H), 7.07 (dt, J=8.7, 1.7 Hz, 1H), 5.07 (dd, J=13.2, 5.0 Hz, 1H), 4.38 (dd, J=17.2, 9.0 Hz, 1H), 4.31-4.16 (m, 2H), 3.53 (dt, J=11.9, 4.7 Hz, 2H), 3.44 (ddd, J=11.8, 9.0, 3.2 Hz, 2H), 2.90 (ddd, J=17.2, 13.6, 5.5 Hz, 1H), 2.65-2.54 (m, 2H), 2.48-2.29 (m, 3H), 2.15-2.02 (m, 1H), 2.02-1.93 (m, 2H), 1.72-1.60 (m, 2H), 1.48-1.08 (m, 8H), 0.87 (s, 3H).

Example 85: 3-(1-oxo-5-(((1S,2S)-2-((pyrimidin-5-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-103)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.16 g, 0.184 mmol) in DMF (2 mL) was added pyrimidine-5-carbaldehyde (85-1, 0.02 mL, 0.26 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (58 mg, 0.28 mmol) was added and stirring was continued at rt for 24 h. Additional pyrimidine-5-carbaldehyde (85-1, 12 mg, 0.11 mmol) was added and the reaction mixture was stirred at rt for 15 min. Additional sodium triacetoxyborohydride (25 mg, 0.12 mmol) was added stirring was continued at rt for 1 h. Additional pyrimidine-5-carbaldehyde (85-1, 12 mg, 0.11 mmol) was added and the reaction mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (25 mg, 0.12 mmol) was the again added and stirring was continued at rt for 1 h. The reaction mixture was then quenched with saturated aqueous sodium bicarbonate and extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N as a modifier) in heptane to afford I-103 (67 mg, 0.14 mmol, 79% yield) as a white solid. MS [M+H]⁺=450.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 9.04 (s, 1H), 8.74 (s, 2H), 7.61 (d, J=8.3 Hz, 1H), 7.20 (s, 1H), 7.08 (d, J=8.5 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.43-4.18 (m, 3H), 3.92-3.76 (m, 1H), 3.14-3.03 (m, 1H), 2.91 (ddd, J=17.9, 13.5, 5.3 Hz, 1H), 2.64-2.55 (m, 1H), 2.44-2.31 (m, 1H), 2.29 (s, 1H), 2.13-1.93 (m, 3H), 1.73-1.59 (m, 2H), 1.42-1.13 (m, 4H).

Example 86: 3-(1-oxo-5-(((1S,2S)-2-((2-(tetrahydro-2H-pyran-4-yl)ethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-104)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.16 g, 0.18 mmol) in DMF (2 ml) was added tetrahydropyranyl-4-acetaldehyde (86-1, 0.034 mL, 0.26 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (58 mg, 0.275 mmol) was added and stirring was continued at rt for 1 h. Additional tetrahydropyranyl-4-acetaldehyde (86-1, 8 μL, 0.06 mmol) was added and the reaction mixture was stirred at rt for 15 minutes. Additional sodium triacetoxyborohydride (25 mg, 0.12 mmol) was added and stirring was continued at rt for 1 h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate and extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N as a modifier) in heptane to afford I-104 (87 mg, 0.18 mmol, 99%) as a white solid. MS [M+H]⁺=470.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.21 (d, J=2.7 Hz, 1H), 7.07 (d, J=8.4 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.38 (dd, J=17.2, 7.1 Hz, 1H), 4.25 (dd, J=17.0, 6.1 Hz, 2H), 3.77 (dd, J=11.3, 4.2 Hz, 2H), 3.23-3.12 (m, 2H), 2.98-2.82 (m, 1H), 2.74-2.55 (m, 4H), 2.38 (qd, J=13.2, 3.9 Hz, 1H), 2.11-2.03 (m, 1H), 2.02-1.92 (m, 2H), 1.72-1.59 (m, 2H), 1.54-1.44 (m, 3H), 1.40-0.98 (m, 8H).

Example 87: 3-(5-(((1S,2S)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-105)

Step 1. Diethyl 2-(3-(2-ethoxy-2-oxoethyl)oxetan-3-yl)malonate (87-3)

A suspension of 60% sodium hydride in mineral oil (4.21 g, 105 mmol) in THF (70 mL), under an atmosphere of nitrogen, was cooled to 0° C. and diethyl malonate (87-2, 18.5 mL, 122 mmol) was added dropwise. The resulting mixture was allowed to warm to rt and stirred at rt for 20 minutes. TBAI (5.17 g, 14.0 mmol) was then added, followed by a solution of ethyl 2-(oxetan-3-ylidene)acetate (87-1, 4.96 g, 34.9 mmol) in THF (8.5 mL) dropwise. The reaction mixture was stirred at rt overnight, cooled to 0° C., quenched with acetic acid (7.5 mL), allowed to warm to room temperature, and stirred at rt for 30 min. The solution was diluted with diethyl ether (250 mL) and the organic phase washed with brine (50 mL), dried over magnesium sulfate, filtered, and concentrated. The crude material was purified via silica gel chromatography eluting with 0-40% EtOAc in heptane to afford 87-3 (7.98 g, 25.1 mmol, 72% yield) as a colorless oil. MS [M+H]⁺=303.3. ¹H NMR (400 MHz, DCM-d₂) δ 4.71 (d, J=6.8 Hz, 2H), 4.52 (d, J≤6.8 Hz, 2H), 4.20 (q, J=7.1 Hz, 4H), 4.11 (q, J=7.1 Hz, 2H), 3.93 (s, 1H), 2.94 (s, 2H), 1.30-1.24 (m, 9H).

Step 2. Diethyl 2,2′-(oxetane-3,3-diyl)diacetate (87-4)

To a solution of diethyl 2-(3-(2-ethoxy-2-oxoethyl)oxetan-3-yl)malonate (87-3, 7.98 g, 26.4 mmol) in DMSO (100 mL) and water (1.5 mL, 83 mmol) was added sodium chloride (3.09 g, 52.8 mmol). The resulting mixture was stirred at 160° C. for 2 h, and cooled to rt and stirred at rt overnight. The reaction mixture was diluted with diethyl ether (300 mL) and washed with brine (2×80 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated to dryness. The crude product was diluted with DMSO (50 mL) and water (1.5 mL, 83 mmol) and sodium chloride (3.09 g, 52.8 mmol) was added. The solution was then stirred at 160° C. for 2 days. The solution was diluted with diethyl ether (300 mL) and washed with brine (80 mL). The organic layer was then dried over magnesium sulfate, filtered, and concentrated. The crude material was purified via silica gel chromatography eluting with 0-50% EtOAc in heptane to afford 87-4 (4.28 g, 18.6 mmol, 70% yield), as a colorless oil. MS [M+H]⁺=231.3. ¹H NMR (400 MHz, DCM-d₂) δ 4.54-4.48 (m, 2H), 4.28-4.16 (m, 2H), 4.16-4.06 (m, 4H), 2.87 (s, 4H), 1.33-1.21 (m, 6H).

Step 3. 2,2′-(oxetane-3,3-diyl)bis(ethan-1-ol) (87-5)

To LiAlH₄ (2.15 g, 56.6 mmol) dissolved in THF (75 mL) under an atmosphere of nitrogen and cooled to 0° C., was added a solution of diethyl 2,2′-(oxetane-3,3-diyl)diacetate (87-4, 4.28 g, 18.6 mmol) in THF (10 mL). The resulting mixture was allowed to slowly warmed to rt and stirred at rt for 1 h. The reaction mixture was cooled to 0° C. and water (4.2 mL) and 1M NaOH (aq) (8.4 mL) were added. The solution was filtered through Celite®, and the Celite® pad was rinsed with EtOAc. The solution was concentrated, diluted with ethyl acetate (150 mL), and washed with brine (40 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness to afford 87-5 (434 mg, 2.97 mmol, 16% yield) a yellow solid, which was carried onto the next step without purification.

Step 4. Oxetane-3,3-diylbis(ethane-2,1-diyl) bis(4-methylbenzenesulfonate) (87-6)

To 2,2′-(oxetane-3,3-diyl)bis(ethan-1-ol) (87-5, 434 mg, 2.97 mmol) and tosyl chloride (1.43 g, 7.52 mmol) dissolved in MeCN (15 mL) was added TEA (1.5 mL, 12 mmol) and DMAP (36 mg, 0.30 mmol) and the resulting mixture was stirred at rt overnight. The solution was then diluted with EtOAc (250 mL) and washed with water (50 mL) and brine (40 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc in heptane to afford 87-6 (170 mg, 026 mmol, 9% yield). MS [M+H]⁺=455.4. ¹H NMR (400 MHz, DCM-d₂) δ 7.83-7.71 (m, 4H), 7.46-7.35 (m, 4H), 4.39-4.33 (m, 2H), 4.29 (s, 2H), 4.05 (t, J=6.4 Hz, 4H), 2.46 (s, 6H), 2.04-1.96 (m, 4H).

Step 5. 3-(5-(((1S,2S)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-105)

3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 101.8 mg, 0.29 mmol) and oxetane-3,3-diylbis(ethane-2,1-diyl) bis(4-methylbenzenesulfonate) (87-6, 169.7 mg, 0.37 mmol) were dissolved in MeCN (2 mL) and DIPEA (0.15 mL, 0.86 mmol) and the resulting mixture was stirred at 120° C. in a μW reactor for 4 h. The reaction mixture was concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N) in DCM (with 0.1% Et₃N) and then purified further via reverse phase preparative HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford the formate salt of I-105 (18 mg, 0.035 mmol, 12% yield) as a white solid. MS [M+H]⁺=468.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.14 (s, 1H), 7.59 (dd, J=8.4, 1.0 Hz, 1H), 7.17 (s, 1H), 7.02 (dt, J=8.4, 2.9 Hz, 1H), 5.06 (dd, J=13.2, 5.1 Hz, 1H), 4.50-4.19 (m, 3H), 4.19-4.04 (m, 4H), 2.90 (ddd, J=18.0, 13.5, 5.3 Hz, 1H), 2.63-2.53 (m, 3H), 2.46-2.34 (m, 3H), 2.16-2.08 (m, 1H), 2.05-1.91 (m, 1H), 1.78-1.45 (m, 7H), 1.42-1.12 (m, 5H).

Example 88: 3-(5-(((1S,2S)-2-(((4-methoxytetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-106)

Step 1. 4-Methoxytetrahydro-2H-pyran-4-carbaldehyde (88-2)

To (4-methoxyoxan-4-yl)methanol (88-1, 0.200 g, 1.37 mmol) dissolved in DCM (6.8 mL) and cooled to 0° C. was added DMP (812 mg, 1.92 mmol) followed by 2 drops of water. The resulting mixture was stirred at 0° C. for 1 h and then at rt for 1 h. The reaction mixture was quenched with 1:1:2 saturated aqueous sodium bicarbonate:saturated aqueous sodium thiosulfate:diethyl ether and stirred until the cloudy mixture turned clear. The organic phase was separated and the aqueous phase was extracted with diethyl ether (×2). The combined organic phases were passed through a phase separator and concentrated to afford 88-2 as a colorless oil, which was taken onto the next reaction without purification.

Step 2. 3-(5-(((1S,2S)-2-(((4-methoxytetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-106)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.10 g, 0.12 mmol) in DMF (1.5 mL) was added 4-methoxytetrahydro-2H-pyran-4-carbaldehyde (88-2, 31 mg, 0.16 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added and stirring was continued at rt for 18 h. The reaction mixture was concentrated down, magnesium sulfate (14 mg, 0.12 mmol) was added, and stirring was continued at 45° C. for 16 h. Additional 4-methoxytetrahydro-2H-pyran-4-carbaldehyde (88-2, 22 mg, 0.12 mmol) was added and the reaction mixture stirred at r.t. for 15 minutes. Sodium triacetoxyborohydride (29 mg, 0.14 mmol) was added and stirring was continued at rt for 1 h. Additional 4-methoxytetrahydro-2H-pyran-4-carbaldehyde (88-2, 22 mg, 0.12 mmol) was added and stirring was continued at rt for 15 mins. Sodium triacetoxyborohydride (29 mg, 0.14 mmol) was then again added. The reaction mixture was stirred at rt for 1 h and then quenched with saturated aqueous sodium bicarbonate and extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separator and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in heptane to afford I-106 (57 mg, 0.11 mmol, 97% yield) as a white solid. MS [M+H]⁺=486.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.21 (s, 1H), 7.07 (dd, J=8.6, 2.2 Hz, 1H), 5.07 (dd, J=13.2, 5.0 Hz, 1H), 4.38 (dd, J=17.2, 11.5 Hz, 1H), 4.32-4.15 (m, 2H), 3.55-3.42 (m, 4H), 3.04 (s, 3H), 2.91 (ddd, J=17.1, 13.5, 5.3 Hz, 1H), 2.69-2.53 (m, 4H), 2.43-2.31 (m, 1H), 2.15-2.04 (m, 1H), 2.01-1.93 (m, 2H), 1.75-1.11 (m, 10H).

Example 89: 3-(5-(((1S,2S)-2-(((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-107)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 100 mg, 0.280 mmol) in DCE (2 mL) was added 2,2-dimethyltetrahydro-2H-pyran-4-carbaldehyde (89-1, 42 mg, 0.29 mmol) and the resulting mixture was stirred at rt for 30 minutes. Sodium triacetoxyborohydride (178 mg, 0.839 mmol) was added and stirring was continued at rt for 2 days. The reaction mixture was then added to a saturated aqueous solution of sodium hydrogen carbonate (20 mL). The aqueous phase was extracted with 20% i-PrOH/DCM (×3) and the combined organic phases were passed through a phase separating column and concentrated. The crude material was purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% Et₃N) in DCM (with 0.1% NEt₃) to afford I-107 (42 mg, 0.083 mmol, 30% yield) as a yellow solid. MS [M+H]⁺=484.4. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm) 10.95 (s, 1H), 7.65-7.55 (m, 1H), 7.20 (d, J=2.3 Hz, 1H), 7.06 (dd, J=8.4, 2.1 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.43-4.18 (m, 3H), 3.58-3.51 (m, 1H), 3.51-3.42 (m, 1H), 2.90 (ddd, J=17.2, 13.6, 5.4 Hz, 1H), 2.68-2.54 (m, 2H), 2.47-2.31 (m, 3H), 2.10-2.01 (m, 1H), 2.01-1.90 (m, 2H), 1.75-1.60 (m, 3H), 1.59-1.11 (m, 6H), 1.11-1.01 (m, 6H), 1.01-0.78 (m, 2H).

Example 90: 3-(5-(((1S,2S)-2-(3-(benzyloxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-54)

A solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 50 mg, 0.14 mmol), (((1,3-dibromopropan-2-yl)oxy)methyl)benzene (120-1, 129 mg, 0.42 mmol), and DIPEA (0.12 mL, 0.70 mmol) in MeCN (1 mL) was stirred at 120° C. under μW irradiation for 3 h. The reaction mixture was then concentrated to dryness, treated with DCM, and washed with saturated NaHCO₃ (aq). The phases were separated and the aqueous phase was extracted with DCM (×3). The combined organic phases were concentrated. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N) in DCM (with 0.1% Et₃N) and then further purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient. The fractions containing the desired product were combined and lyophilized to afford I-54 (7 mg, 0.012 mmol, 9% yield) as a white solid. MS [M+H]⁺=504.5. ¹H NMR (400 MHz, DCM-d₂) δ 8.21 (br s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.28-7.16 (m, 5H), 6.93 (s, 1H), 6.91-6.85 (m, 1H), 5.10-4.94 (m, 1H), 4.31 (s, 2H), 4.29-4.16 (m, 3H), 4.12-4.02 (m, 1H), 3.61 (s, 2H), 3.31-2.93 (m, 2H), 2.84-2.65 (m, 2H), 2.58-2.34 (m, 1H), 2.33-2.17 (m, 1H), 2.12-2.04 (m, 1H), 1.97 (s, 1H), 1.81 (s, 1H), 1.64 (s, 2H), 1.36-1.05 (m, 4H).

Example 91: 3-(1-oxo-5-(((1S,2S)-2-(3-(pyridazin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-204)

Step 1. 3-chloro-6-((2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilaundecan-6-yl)oxy)pyridazine (182-2)

To a solution of 2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilaundecan-6-ol (116-2, 2.00 g, 6.25 mmol) in THF (50 mL) was added NaH (0.370 g, 9.37 mmol) portion wise at 0° C. and the resulting mixture was stirred at rt for 1 h. The reaction mixture was cooled to 0° C., 3,6-dichloropyridazine (182-1, 0.920 g, 6.25 mmol) was added and was then refluxed for 16 h. The reaction mixture was quenched with cold water and extracted with EtOAc (3×50 mL). The combined organic phases were washed with water, brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 10% EtOAc in hexanes to afford 182-2 (2.20 g, 5.08 mmol, 81% yield) as colorless liquid. ¹H NMR (400 MHz, CDCl₃): δ 7.35 (d, J=9.2 z, 1H), 6.95-6.94 (m, 1H), 3.94-3.91 (m, 4H), 3.65-3.64 (m, 1H), 0.86 (s, 18H), 0.02 (s, 12H).

Step 2. 3-((2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilaundecan-6-yl)oxy)pyridazine (182-3)

To a flask containing a deoxygenated solution of 182-2 (2.20 g, 5.03 mmol) in EtOAc-THF (1:1) (40 mL) under inert atmosphere was added Et₃N (2.2 mL, 15 mmol) and 10% Pd—C (0.55 g, 25% wt/wt) and the flask was purged with hydrogen and then stirred under a hydrogen atmosphere using a balloon at rt for 2 h. The reaction mixture was filtered through Celite®, the pad was washed with EtOAc, and the filtrate was concentrated to dryness to afford crude 182-3 (2.0 g), as pale yellow liquid. The crude material was carried on to the next step without purification. MS [M+H]⁺=399.3.

Step 3. 2-(pyridazin-3-yloxy)propane-1,3-diol (182-5)

To a stirred solution of 182-3 (2.00 g, 5.02 mmol) in MeOH (30 mL) was added concentrated HCl (4.0 mL) drop wise at 0° C. and the resulting mixture was stirred at room temperature for 4 h. The reaction mixture was concentrated to dryness. The obtained crude material was diluted with cooled water, basified with NaHCO₃ and concentrated to dryness. The residue obtained was stirred with 10% MeOH in DCM, filtered and the filtrate was concentrated to dryness to afford crude 182-4 (0.60 g), as pale brown liquid. The crude material was carried on to the next step without purification. ¹H NMR (300 MHz, CDCl₃): δ 8.38 (d, J=4.2 Hz, 1H), 7.73-7.39 (m, 1H), 7.06 (d, J=4.2 Hz, 1H), 4.68-4.63 (m, 2H), 4.16-4.13 (m, 1H), 3.81-3.68 (m, 2H).

Step 4. 2-(pyridazin-3-yloxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (182-5)

To a solution of 182-4 (0.60 g, 3.50 mmol) in DCM (30 mL) was added Et₃N (2.02 mL, 14.10 mmol), DMAP (0.017 g, 0.14 mmol) and TsCl (1.40 g, 7.70 mmol) at 0° C. and the resulting mixture was stirred at rt for 3 h. The reaction mixture was quenched with cooled water and extracted with DCM (3×30 mL). The combined organic extracts were washed with water, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 50% EtOAc in hexanes to afford 182-5 (0.80 g, 1.67 mmol, 50% yield) as colorless liquid. MS [M+H]⁺=479.1.

Step 5. 3-(1-oxo-5-(((1S,2S)-2-(3-(pyridazin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-204)

To a solution of I-15 (200 mg, 0.56 mmol) in MeCN (5 mL) was added 182-5 (168 mg, 0.66 mmol) and DIPEA (0.72 mL, 3.36 mmol) and the resulting mixture was stirred at 120° C. for 2 h. The reaction mixture was then diluted with sat. sodium bicarbonate solution and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified by reverse phase HPLC (Column: LUNA C18 (250 mm×21.2 mm), 5.0 g, Mobile phase A: 0.01% HCOOH (aq), Mobile phase B: MeCN, Method: 0-75% MeCN over 6 minutes. Flow rate: 20 ml/min) to afford the formate salt of I-204 (12 mg, 0.02 mmol, 5% yield), as a white solid. MS [M+H]⁺=492.4. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 8.01-7.99 (m, 1H), 7.90-7.89 (m, 1H), 7.57 (d, J=8.4 Hz, 2H), 7.17 (s, 1H), 7.05 (d, J=8.4 Hz, 1H), 6.93-6.90 (m, 1H), 6.35-6.34 (m, 1H), 5.31-5.26 (m, 1H), 5.08-5.04 (m, 1H), 4.30-4.23 (m, 4H), 3.85-3.75 (m, 1H), 2.91-2.89 (m, 2H), 2.66-2.60 (m, 1H), 2.36-2.32 (m, 1H), 2.07-1.96 (m, 3H), 1.90-1.85 (m, 1H), 1.70-1.65 (m, 2H), 1.37-1.31 (m, 4H).

Example 92: 3-(5-(((1S,2S)-2-((7-oxaspiro[3.5]nonan-2-yl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-110)

Step 1. 7-oxaspiro[3.5]nonan-2-yl 4-methylbenzenesulfonate (92-2)

TsCl (174 mg, 0.914 mmol) was added to a solution of 7-oxaspiro[3.5]nonan-2-ol (92-1, 100 mg, 0.703 mmol), DMAP (9 mg, 0.07 mmol) and TEA (0.20 mL, 1.4 mmol) in MeCN (3 mL). The resulting mixture was stirred at rt for 16 h and then concentrated to dryness. The crude material was treated with diethyl ether and the obtained solid was rinsed with diethyl ether several times. The filtrate was collected and concentrated to dryness to afford crude 92-2 (240 mg) as a gold colored oil. ¹H NMR (400 MHz, Chloroform-d) δ 7.80-7.75 (m, 2H), 7.36-7.32 (m, 2H), 4.88-4.77 (m, 1H), 3.59-3.50 (m, 4H), 2.46 (s, 3H), 2.31-2.21 (m, 2H), 1.98-1.89 (m, 2H), 1.58-1.55 (m, 2H), 1.53-1.49 (m, 2H).

Step 2. 3-(5-(((1S,2S)-2-((7-oxaspiro[3.5]nonan-2-yl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-110)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 100 mg, 0.280 mmol) and crude 7-oxaspiro[3.5]nonan-2-yl 4-methylbenzenesulfonate (92-2, 134 mg, 0.420 mmol) in MeCN (1 mL) was added DIPEA (0.15 mL, 0.84 mmol) and the resulting mixture was stirred at 160° C. under μW irradiation for 18 h. The reaction mixture was concentrated, taken up in DCM (10 mL), and extracted with 1M HCl (aq) (3×10 mL). The combined aqueous phases were washed with DCM (3×10 mL). The phases were separated and the aqueous phase was cooled and basified to a pH of 9-10 using solid NaHCO₃. The basic layer was extracted with DCM (4×20 mL) and the combined organic phases were dried over Na₂SO₄, filtered, and concentrated. The crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collection tubes contained several drops of formic acid) and then treated with DCM and washed with saturated aqueous NaHCO₃. The phases were separated and the organic phase was concentrated to dryness to afford I-110 (6 mg, 0.01 mmol, 4% yield), as a white solid. MS [M+H]⁺=482.4. ¹H NMR (400 MHz, DCM-d₂) δ 7.62 (dd, J=8.1, 0.9 Hz, 1H), 6.96-6.91 (m, 2H), 5.07-4.98 (m, 1H), 4.32-4.17 (m, 2H), 4.04-3.96 (m, 1H), 3.52-3.35 (m, 4H), 3.28 (quint, J=7.7 Hz, 1H), 2.78-2.57 (m, 3H), 2.32-2.16 (m, 1H), 2.16-1.99 (m, 4H), 1.97-1.88 (m, 1H), 1.68-1.57 (m, 2H), 1.50-0.99 (m, 11H)

Example 93: 1-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)cyclobutane-1-carbonitrile (I-111)

Step 1. 1-(Hydroxymethyl)cyclobutane-1-carbonitrile (93-2)

To a solution of 1-cyanocyclobutane-1-carboxylic acid (93-1, 450 mg, 3.6 mmol) in THF (18 mL) at 0° C. was added 1M BH₃ in THF (11 mL, 11 mmol). The resulting mixture was allowed to warm slowly to rt and then stirred at rt for 18 h. MeOH was then slowly, followed by aqueous saturated aqueous bicarbonate solution until neutralized. The aqueous solution was extracted with DCM, the phases were separated, and the organic phase was passed through a phase separating column and concentrated to afford 93-2 (540 mg, 3.55 mmol, 99% yield) as a crude oil which was taken onto the next step without purification. ¹H NMR (400 MHz, DMSO-d₆) δ 4.31 (t, J=5.2 Hz, 1H), 3.35-3.29 (m, 2H), 1.54-1.47 (m, 2H), 1.45-1.24 (m, 4H).

Step 2. 1-Formylcyclobutane-1-carbonitrile (93-3)

To a solution of 1-(hydroxymethyl)cyclobutane-1-carbonitrile (93-2, 360 mg, 2.34 mmol) in DCM (6 mL) was added DMP (1.50 g, 3.55 mmol) and the resulting mixture was stirred at rt for 4 h. Additional DMP (280 mg, 0.66 mmol) was added and stirring was continued at rt for 16 h. The reaction mixture was diluted with 20% sodium thiosulfate (aq) and extracted with EtOAc (×2). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to afford 93-3 (272 mg, 2.34 mmol, quantitative yield) as a crude orange yellow oil that was used in the next step without purification.

Step 3. 1-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)cyclobutane-1-carbonitrile (I-111)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 200 mg, 0.560 mmol) in TFE (3 mL) was added 1-formylcyclobutane-1l-carbonitrile (93-3, 183 mg, 1.68 mmol, in DCM (3 mL) followed by sodium triacetoxyborohydride (356 mg, 1.68 mmol). The resulting mixture was stirred at rt for 3 days. Additional sodium triacetoxyborohydride (225 mg, 1.06 mmol) was added and stirring was continued at 40° C. and then at rt. The reaction mixture was filtered over Celite® eluting with DCM and the filtrate was concentrated to dryness. The crude material was purified via C-18 reverse phase chromatography eluting with 10-100% MeCN/water with 0.1% formic acid and the further purified using reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collection tubes contained several drops of formic acid). The fraction containing the desired product were combined and lyophilized to afford I-111 (0.7 mg, 1 μmol, 0.2% yield). MS [M+H]⁺=451.2. ¹H NMR (400 MHz, DCM-d₂) δ 7.95 (s, 1H), 7.71 (d, J=8.5 Hz, 1H), 7.04 (d, J=5.7 Hz, 2H), 5.13 (dt, J=13.5, 4.3 Hz, 1H), 4.48-4.23 (m, 2H), 4.17-4.10 (m, 1H), 2.95 (s, 2H), 2.89-2.73 (m, 3H), 2.47-2.31 (m, 2H), 2.24-1.91 (m, 7H), 1.77 (m, 4H), 1.44-1.22 (m, 4H).

Example 94: 3-(5-(((1S,2S)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-112)

Step 1. 2-(2-chlorophenoxy)propane-1,3-diol (94-2)

Sodium borohydride (189 mg, 5.00 mmol) was added to a solution of dimethyl 2-(2-chlorophenoxy)malonate (94-1, 259 mg, 1.0 mmol) in THF (4 mL) and MeOH (1.0 mL). The resulting mixture was stirred at rt for 90 minutes and then quenched with water. The phases were separated and the aqueous phase was extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness to afford 94-2 (105 mg, 0.518 mmol, 52% yield) as a colorless oil. ¹H NMR (400 MHz, Chloroform-d) δ 7.20-7.15 (m, 2H), 6.89-6.81 (m, 2H), 4.31 (quint, J=4.8 Hz, 1H), 3.89-3.76 (m, 4H).

Step 2. 2-(2-chlorophenoxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (94-3)

TsCl (350 mg, 1.84 mmol) was added to a solution of 2-(2-chlorophenoxy)propane-1,3-diol (94-2, 105 mg, 0.52 mmol), DMAP (6 mg, 0.05 mmol), and TEA (0.3 mL, 2 mmol) in MeCN (2 mL). The resulting mixture was stirred at rt for 4 h, and then filtered and concentrated. The crude material was purified via silica gel chromatography eluting with 0-60% EtOAc in heptane to afford 94-3 (160 mg, 0.313 mmol, 60% yield) as a yellow solid. MS [M+H₂O]⁺=528.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.75-7.70 (m, 4H), 7.34-7.29 (m, 4H), 7.17-7.13 (m, 2H), 6.68-6.63 (m, 2H), 4.52 (quint, J=5.0 Hz, 1H), 4.22-4.12 (m, 4H), 2.45 (s, 6H).

Step 3. 3-(5-(((1S,2S)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-112)

DIPEA (0.10 mL, 0.57 mmol) was added to a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 50 mg, 0.14 mmol) and 2-(2-chlorophenoxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (94-3, 71.5 mg, 0.14 mmol) in MeCN (0.75 mL). The resulting mixture was stirred at 120° C. under μW irradiation for 4 h and then was concentrated. The crude material was purified via silica gel chromatography eluting with 0-50% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N) in DCM (with 0.1% Et₃N) to afford I-112 (30 mg, 0.054 mmol, 39% yield) as a white solid. MS [M+H]⁺=524.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.32-7.26 (m, 2H), 7.17 (s, 1H), 7.03 (d, J=8.2 Hz, 1H), 6.88-6.80 (m, 2H), 5.06 (dd, J=13.2, 5.1 Hz, 1H), 4.70 (quint, J=5.6 Hz, 1H), 4.38 (dd, J=17.2, 7.0 Hz, 1H), 4.32-4.21 (m, 2H), 3.76-3.64 (m, 2H), 3.22-3.14 (m, 1H), 3.09-3.01 (m, 1H), 2.95-2.85 (m, 1H), 2.64-2.55 (m, 1H), 2.44-2.34 (m, 2H), 2.04-1.94 (m, 2H), 1.89-1.79 (m, 1H), 1.71-1.61 (m, 2H), 1.42-1.32 (m, 2H), 1.31-1.05 (m, 2H).

Example 95: 3-(5-(((1S,2S)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-113)

Step 1. 2-(2-methoxyphenoxy)propane-1,3-diol (95-2)

Sodium borohydride (189 mg, 5.00 mmol) was added to a solution of dimethyl 2-(2-methoxyphenoxy)malonate (95-1, 254 mg, 1 mmol) in THF (4 mL) and MeOH (1 mL). The resulting mixture was stirred at rt for 90 minutes and then quenched with water. The layers separated and the aqueous phase was extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated to afford 95-2 (103 mg, 0.520 mmol, 52% yield), which was taken onto the next step without purification.

Step 2. 2-(2-methoxyphenoxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (95-3)

TsCl (248 mg, 1.30 mmol) was added to a solution of 2-(2-methoxyphenoxy)propane-1,3-diol (95-2, 103 mg, 0.520 mmol), DMAP (6 mg, 0.05 mmol), and TEA (0.3 mL, 2 mmol) in MeCN (2 mL). The resulting mixture was stirred at rt for 4 h and then concentrated. The crude material was purified via silica gel chromatography eluting with 0-60% EtOAc in heptane to afford 95-3 (100 mg, 0.20 mmol, 38% yield) as a light brown oil. MS [M+H]⁺=507.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.78-7.72 (m, 4H), 7.35-7.30 (m, 4H), 7.01 (ddd, J=8.2, 6.6, 2.5 Hz, 1H), 6.85 (dd, J=8.1, 1.3 Hz, 1H), 6.81-6.74 (m, 2H), 4.44 (quint, J=5.0 Hz, 1H), 4.26-4.15 (m, 4H), 3.76 (s, 3H), 2.45 (s, 6H)

Step 3. 3-(5-(((1S,2S)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-113)

DIPEA (0.05 mL, 0.3 mmol) was added to a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 35.3 mg, 0.1 mmol) and 2-(2-methoxyphenoxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (95-3, 50 mg, 0.1 mmol) in MeCN (0.75 mL). The resulting mixture was stirred at 120° C. under μW irradiation for 8 h and then concentrated. The crude material was purified via silica gel chromatography eluting with 0-65% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N) in DCM (with 0.1% Et₃N) to afford I-113 (20 mg, 0.037 mmol, 37% yield) as a white solid. MS [M+H]⁺=520.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.2 Hz, 1H), 7.04 (dd, J=8.5, 2.2 Hz, 1H), 6.95 (dd, J=8.0, 1.6 Hz, 1H), 6.88 (td, J=7.7, 1.6 Hz, 1H), 6.81 (td, J=7.7, 1.7 Hz, 1H), 6.70 (dd, J=7.9, 1.6 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.69-4.59 (m, 1H), 4.38 (dd, J=17.2, 6.4 Hz, 1H), 4.26 (dd, J=17.1, 5.2 Hz, 2H), 3.76-3.64 (m, 2H), 3.74 (s, 3H), 3.25-3.17 (m, 1H), 3.05 (t, J=6.4 Hz, 1H), 2.97-2.84 (m, 1H), 2.64-2.55 (m, 1H), 2.45-2.33 (m, 2H), 2.05-1.93 (m, 2H), 1.90-1.78 (m, 1H), 1.70-1.59 (m, 2H), 1.42-1.05 (m, 4H).

Example 96: 3-(1-oxo-5-(((1S,2S)-2-((pyrazolo[1,5-a]pyrimidin-6-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-114)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.10 g, 0.28 mmol) in DMF (0.93 mL), was added pyrazolo[1,5-a]pyrimidine-6-carbaldehyde (96-1, 49 mg, 0.34 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (89 mg, 0.42 mmol) was added and stirring was continued at rt for 16 h. The reaction mixture was concentrated and then diluted with DCM (1 mL) and MeCN (2 mL). Additional pyrazolo[1,5-a]pyrimidine-6-carbaldehyde (96-1, 20 mg, 0.14 mmol) was added and the reaction mixture was sonicated for 30 minutes at rt. Sodium triacetoxyborohydride (50 mg, 0.24 mmol) was added and the resulting mixture was sonicated for another 30 minutes at rt. The reaction mixture was concentrated onto Celite® and the crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N) in heptane and then further purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-114 (9 mg, 0.02 mmol, 6% yield) as a white solid. MS [M+H]⁺=489.4 ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 9.00-8.95 (m, 1H), 8.53 (t, J=2.2 Hz, 1H), 8.14 (d, J=2.1 Hz, 1H), 7.58 (dd, J=8.4, 2.0 Hz, 1H), 7.17 (dd, J=4.3, 2.1 Hz, 1H), 7.07 (ddd, J=8.4, 3.6, 2.2 Hz, 1H), 6.66 (d, J=2.4 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.39-4.15 (m, 3H), 3.89 (q, J=14.3 Hz, 2H), 2.91 (ddd, J=17.1, 13.5, 5.4 Hz, 1H), 2.71-2.55 (m, 2H), 2.44-2.35 (m, 1H), 2.13-1.92 (m, 3H), 1.65 (d, J=10.1 Hz, 2H), 1.42-1.23 (m, 4H).

Example 97: 3-(5-(((1S,2S)-2-((4,4-difluorocyclohexyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-115)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 50 mg, 0.14 mmol), 4,4-difluorocyclohexan-1-one (97-1, 375 mg, 2.8 mmol), and MgSO₄ (51 mg, 0.42 mmol) in DMF (0.5 mL) was added with NaBH(OAc)₃ (59 mg, 0.28 mmol) in one portion. The resulting mixture was stirred at 60° C. for 16 h and then diluted with DCM (2 mL), filtered, and concentrated. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM to afford I-115 (36 mg, 0.08 mmol, 55% yield) as a white solid. MS [M+H]⁺=476.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.69 (d, J=8.4 Hz, 1H), 7.10-6.86 (m, 2H), 5.10 (ddd, J=13.3, 5.2, 2.4 Hz, 1H), 4.43-4.21 (m, 2H), 4.13-3.96 (m, 1H), 2.93-2.70 (m, 4H), 2.31 (qd, J=12.9, 5.9 Hz, 1H), 2.21-2.09 (m, 2H), 2.07-1.89 (m, 5H), 1.88-1.64 (m, 6H), 1.42-1.24 (m, 5H).

Example 98: 3-(5-(((1S,2S)-2-((2,4-difluorobenzyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-116)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 100 mg, 0.28 mmol) in DMF (2 mL) under an atmosphere of nitrogen was added 2,4-difluorobenzaldehyde (98-1, 0.03 mL, 0.31 mmol) and the resulting mixture was stirred at rt for 30 minutes. Sodium triacetoxyborohydride (178 mg, 0.839 mmol) was then added and stirring was continued at rt for 16 h. The reaction mixture was added to a saturated aqueous solution of sodium hydrogen carbonate (30 mL). The resulting suspension was sonicated and filtered and the solid was rinsed with water (×3). The filtrate was extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and concentrated. The isolated solid and product from extraction were combined and purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-116 (22 mg, 0.04 mmol, 15% yield) as a white solid. MS [M+H]⁺ 484.2. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.48 (td, J=8.7, 6.8 Hz, 1H), 7.20-7.10 (m, 2H), 7.08-6.98 (m, 2H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.42-4.19 (m, 3H), 3.87-3.73 (m, 2H), 2.90 (ddd, J=18.0, 13.6, 5.4 Hz, 1H), 2.68-2.55 (m, 2H), 2.39 (td, J=13.0, 4.3 Hz, 1H), 2.10-2.02 (m, 1H), 2.02-1.93 (m, 2H), 1.70-1.61 (m, 2H), 1.41-1.16 (m, 4H).

Example 99: 3-(5-(((1S,2S)-2-(3-isopropoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-53)

Step 1. 3-(5-(((1S,2S)-2-(3-hydroxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (99-2)

Epichlorohydrin (99-1, 0.12 mL, 1.5 mmol) and DIPEA (0.12 mL, 0.70 mmol) were added to a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 50 mg, 0.14 mmol) in MeCN (4 mL) and the resulting mixture was stirred at 120° C. under μW irradiation for 1 h, then stirred at 130° C. under μW irradiation for 1 h. The reaction mixture was concentrated, taken up in DCM, and extracted with 1M HCl (aq) (×3). The combined aqueous phases were washed with DCM. The phases were separated and the aqueous phase was cooled and basified to a pH of 9-10 with solid NaHCO₃. The basic aqueous phase was extracted with DCM (×4) and the combined organic phases were dried over Na₂SO₄, filtered, and concentrated to afford crude 99-2 (80 mg) as a pale yellow solid. MS [M+H]⁺=414.3, which was taken onto the next step without purification.

Step 2. 3-(5-(((1S,2S)-2-(3-isopropoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-53)

2-Bromopropane (165-1, 0.16 mL, 1.7 mmol) was added to a solution of 3-(5-(((1S,2S)-2-(3-hydroxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (99-2, 70 mg, 0.17 mmol) and AgOTf (217 mg, 0.85 mmol) in DCM (0.5 mL) and the resulting mixture was stirred at rt for 16 h. The reaction mixture was diluted with DCM and filtered. The filtrate was washed with saturated NaHCO₃ (aq) and the phases were separated. The aqueous phase was extracted with DCM and the combined organic layers were dried over Na₂SO₃, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-53 (9 mg, 0.02 mmol, 11% yield) as a pale yellow solid. MS [M+H]⁺=456.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.15 (s, 1H), 7.02 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.38 (dd, J=17.2, 5.8 Hz, 1H), 4.28-4.18 (m, 2H), 4.01-3.91 (m, 1H), 3.54-3.42 (m, 3H), 2.98-2.84 (m, 2H), 2.77 (t, J=6.4 Hz, 1H), 2.63-2.55 (m, 1H), 2.42-2.35 (m, 1H), 2.31-2.24 (m, 1H), 2.02-1.91 (m, 2H), 1.78 (d, J=12.3 Hz, 1H), 1.68-1.55 (m, 2H), 1.40-1.19 (m 4H), 1.02 (d, J=6.1 Hz, 6H).

Example 100: 3-(5-(((1S,2S)-2-(((1R,2R)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1S,2S)-2-(((1S,2S)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-118 & I-119)

Step 1. (1S,2S)-2-methoxycyclopentyl 4-nitrobenzenesulfonate (100-3)

To (1S,2S)-2-methoxycyclopentan-1-ol (100-1, 110 mg, 0.95 mmol) and 1-methyl-1H-imidazole (0.38 mL, 4.7 mmol) in DCM (10 mL) was added 4-nitrobenzenesulfonyl chloride (100-2, 210 mg, 0.95 mmol). The resulting mixture was stirred overnight at rt and then quenched with saturated aqueous NaHCO₃ and extracted with DCM (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to afford 100-3 (128 mg, 0.43 mmol, 45% yield). The crude product was taken onto the next step without purification.

Step 2. 3-(5-(((1S,2S)-2-(((1R,2R)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione or 3-(5-(((1S,2S)-2-(((1S,2S)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-118 & I-119)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 50 mg, 0.14 mmol) in DIPEA (0.15 mL, 0.84 mmol) and MeCN (0.5 mL) was added (1S,2S)-2-methoxycyclopentyl 4-nitrobenzenesulfonate (100-3, 105 mg, 0.35 mmol) and the resulting mixture was stirred at 120° C. for 4 h. The reaction mixture was concentrated and the crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collected fractions contained ˜3 drops of formic acid). The fractions containing the desired product were combined and lyophilized to afford one diastereomer (I-118, 4.5 mg, 8.6 μmol, 6% yield) as a white solid. MS [M+H]⁺=456.3. ¹H NMR (400 MHz, MeCN-d₃) δ 7.62 (d, J=8.4 Hz, 1H), 7.17-7.10 (m, 1H), 7.03 (dd, J=8.4, 2.2 Hz, 1H), 5.03 (ddd, J=13.4, 5.2, 2.5 Hz, 1H), 4.34 (d, J=16.7 Hz, 1H), 4.26 (dd, J=16.7, 3.8 Hz, 1H), 4.12 (s, 1H), 3.51-3.33 (m, 1H), 3.22 (d, J=2.2 Hz, 3H), 3.14-3.05 (m, 1H), 2.89-2.66 (m, 3H), 2.39 (qd, J=13.2, 4.9 Hz, 1H), 2.12-2.04 (m, 4H), 1.91-1.84 (m, 1H), 1.80-1.67 (m, 3H), 1.64-1.46 (m, 3H), 1.42-1.35 (m, 2H), 1.32-1.26 (m, 2H). The relative stereochemistry of the substituents on the cyclopentane ring was not determined and was assigned arbitrarily.

A second impure product was isolated which was further purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collected fractions contained ˜3 drops of formic acid) to afford the formate salt of the second diastereomer (I-119, 4.4 mg, 8.8 μmol, 6% yield) as a white solid. MS [M+H]⁺=456.3. ¹H NMR (400 MHz, MeCN-d₃+ drop of D₂O) δ 8.29 (s, 1H), 7.60 (dd, J=8.4, 2.9 Hz, 1H), 7.07 (d, J=2.2 Hz, 1H), 7.03-6.94 (m, 1H), 4.96 (ddd, J=13.4, 5.2, 1.4 Hz, 1H), 4.35-4.17 (m, 3H), 3.71-3.54 (m, 1H), 3.27 (s, 2H), 3.25-3.17 (m, 1H), 3.10 (d, J=0.9 Hz, 1H), 2.91-2.81 (m, 1H), 2.80-2.72 (m, 1H), 2.67-2.59 (m, 1H), 2.32 (qd, J=13.2, 4.9 Hz, 1H), 2.17-2.00 (m, 3H), 1.82-1.44 (m, 7H), 1.42-1.16 (m, 5H). The relative stereochemistry of the substituents on the cyclopentane ring was not determined and was assigned arbitrarily.

Example 101: 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)bicyclo[1.1.1]pentane-1-carbonitrile (I-120) Step 1. 3-formylbicyclo[1.1.1]pentane-1-carbonitrile (101-2)

3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carbonitrile (101-1, 124 mg, 1.01 mmol) was dissolved in DCM (5.0 mL) and cooled to 0° C. DMP (540 mg, 1.27 mmol) was added followed by 2 drops of water. The resulting mixture was stirred at rt for 2 h, quenched with saturated aqueous sodium bicarbonate:saturated aqueous sodium thiosulfate:diethyl ether (v/v/v=1:1:2) and then stirred until the cloudy mixture turned clear. The organic phase was separated and the aqueous phase was extracted with diethyl ether (×2). The combined organic phases were passed through a phase separator and concentrated. The crude material was dissolved in DCM (5.0 mL) and cooled to 0° C. DMP (780 mg, 1.84 mmol) was added followed by 3 drops of water. The resulting mixture was stirred at rt for 18 h, quenched with saturated aqueous sodium bicarbonate:saturated aqueous sodium thiosulfate:diethyl ether (v/v/v=1:1:2), and then stirred until the cloudy mixture turned clear. The organic phase was separated and the aqueous phase was extracted with diethyl ether (×2). The combined organic phases were passed through a phase separator and concentrated to afford crude 101-2 as a cream-colored oil. The crude material was taken onto the next step without purification.

Step 2. 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)bicyclo[1.1.1]pentane-1-carbonitrile (I-120)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.15 g, 0.42 mmol) in DMF (1 mL) was added 3-formylbicyclo[1.1.1]pentane-1-carbonitrile (101-2, 61 mg, 0.50 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (133 mg, 0.630 mmol) was added and stirring was continued at rt for 1.5 h. The reaction mixture was then quenched with saturated aqueous sodium bicarbonate and extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separator and concentrated onto Celite®. The crude product was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in heptane to afford I-120 (67.7 mg, 0.14 mmol, 34% yield) as a cream-colored solid. MS [M+H]⁺=463.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.20 (d, J=2.2 Hz, 1H), 7.07 (dd, J=8.2, 2.3 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.38 (dd, J=17.1, 8.5 Hz, 1H), 4.32-4.14 (m, 2H), 2.91 (ddd, J=17.1, 13.6, 5.3 Hz, 1H), 2.70-2.55 (m, 4H), 2.40 (td, J=13.1, 4.5 Hz, 1H), 2.11-2.02 (br s, 6H), 2.02-1.94 (m, 1H), 1.94-1.85 (m, 1H), 1.70-1.59 (m, 2H), 1.39-1.08 (m, 5H).

Example 102: 3-(5-(((1S,2S)-2-(3-(3-fluorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-121)

To a solution of di-tert-butyl (E)-diazene-1,2-dicarboxylate (16.7 mg, 0.073 mmol) in THF (0.25 mL) was added to a solution of 3-(5-(((1S,2S)-2-(3-hydroxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (99-2, 20 mg, 0.048 mmol), triphenylphosphine (19 mg, 0.073 mmol), DIPEA (0.013 mL, 0.073 mmol) and 3-fluorophenol (102-1, 4 μL, 0.05 mmol) in THF (0.25 mL). The resulting mixture was stirred at rt for 16 h and then diluted with DCM and washed with 50% saturated Na₂CO₃ (aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic layers were dried and concentrated. The crude material was purified via reverse phase HPLC (Column: XBridge C18 OBD 30×50 mm; 15-40% MeCN in H₂O+0.1% Formic Acid over 3.5 min at 75 mL/min). The fractions containing the desired product were combined and lyophilized to afford I-121 (8 mg, 0.01 mmol, 27% yield) as a white solid. MS [M+H]⁺=508.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.28 (q, J=8.2 Hz, 1H), 7.21-7.13 (m, 2H), 7.04 (dd, J=8.4, 2.2 Hz, 1H), 6.79-6.72 (m, 1H), 6.71-6.63 (m, 2H), 6.62-6.53 (m, 2H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.78-4.71 (m, 1H), 4.38 (dd, J=17.2, 6.8 Hz, 1H), 4.25 (dd, J=17.2, 5.5 Hz, 2H), 3.79-3.65 (m, 1H), 3.22-3.12 (m, 1H), 3.05 (t, J=6.5 Hz, 1H), 2.95-2.85 (m, 1H), 2.63-2.53 (m, 1H), 2.04-1.92 (m, 2H), 1.88-1.77 (m, 1H), 1.69-1.54 (m, 2H), 1.45-1.22 (m, 4H).

Example 103: 3-(5-(((1S,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-122)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 200 mg, 0.56 mmol) in MeCN (10 mL) was added (3,3-difluorocyclobutyl)methyl 4-methylbenzenesulfonate (53-2, 262 mg, 0.95 mmol) and DIPEA (0.49 mL, 2.8 mmol) and the resulting mixture was stirred at 120° C. for 16 h under μW irradiation. The reaction mixture was then diluted with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated. The crude material was purified via silica gel chromatography eluting with 5% MeOH in DCM to afford I-122 (80 mg, 0.17 mmol, 31% yield) as an off-white solid. MS [M+H]⁺=462.5. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.197 (s, 1H), 7.06 (d, J=8.4 Hz, 1H), 5.08 (dd, J=13.2, 4.8 Hz, 1H), 4.40-4.23 (m, 4H), 2.95-2.86 (m, 1H), 2.66-2.61 (m, 3H), 2.56-2.33 (m, 2H), 2.19-2.16 (m, 3H), 2.08-1.92 (m, 3H), 1.75-1.64 (m, 3H), 1.35-1.14 (m, 5H).

Example 104: 3-(5-(((1S,2S)-2-(((4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-123)

Step 1. cis-Methyl 4-methoxycyclohexane-1-carboxylate (104-2)

To a solution of cis-4-hydroxycyclohexane-1-carboxylic acid (104-1, 2.00 g, 13.9 mmol) in DMF (20 mL) was added NaH (1.38 g, 34.7 mmol) in small portions at 0° C. and the resulting mixture was stirred for 30 min. Methyl iodide (2.6 mL, 42 mmol) was added and stirring was continued at rt for 4 h. Upon complete consumption of the starting material, the reaction mixture was quenched with ice-cold water and extracted with EtOAc (2×100 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 20% EtOAc in heptanes to afford compound 104-2 (1.20 g, 6.97 mmol, 52% yield) as an oil. ¹H NMR (600 MHz, CDCl₃): δ 3.65 (s, 3H), 3.36-3.32 (m, 1H), 3.28 (s, 3H), 2.36-2.32 (m, 1H), 1.88-1.79 (m, 4H), 1.65-1.61 (m, 2H), 1.58-1.47 (m, 2H).

Step 2. cis-4-Methoxycyclohexane-1-carbaldehyde (104-3)

To a solution of cis-methyl 4-methoxycyclohexane-1-carboxylate (104-2, 1.2 g, 6.97 mmol) in THF (15 mL) was added LiAlH₄ (387 mg, 10.46 mmol) dropwise at 0° C. and the resulting mixture was warmed to rt and stirred for 4 h. Upon complete consumption of the starting material, the reaction mixture was quenched with ice-cold water and extracted with EtOAc (3×50 mL). The organic phase was washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. This material was taken in DCM (10 mL), DMP (1.30 g, 6.25 mmol) was added and the resulting mixture was stirred at rt for 4 h. Upon complete consumption of the starting material, the reaction mixture was diluted with DCM (10 mL), filtered through a small pad of Celite® and washed with DCM (10 mL). The combined filtrate was dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography eluting with 10% EtOAc in hexanes to afford 104-3 (450 mg, 3.17 mmol, 76% yield) as colorless liquid. ¹H NMR (300 MHz, CDCl₃): δ 9.61 (s, 1H), 3.37-3.35 (m, 1H), 3.30 (s, 3H), 2.29-2.21 (m, 1H), 1.82-1.57 (m, 8H).

Step 3. 3-(5-(((1S,2S)-2-(((4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-123)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 0.20 g, 0.56 mmol) and cis-4-methoxycyclohexane-1-carbaldehyde (104-3, 0.12 g, 0.84 mmol) in DCE-DMF (9:1) (10 mL) was added NaBH(OAc)₃ (0.18 g, 0.84 mmol) at 0° C. and the resulting mixture was stirred at rt for 16 h. The reaction mixture was then quenched with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified via silica gel chromatography eluting with 5% MeOH in DCM to afford I-123 (50 mg, 0.10 mmol, 18% yield). MS [M+H]⁺=484.5. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.197 (s, 1H), 7.06 (d, J=8.4 Hz, 1H), 5.08 (dd, J=13.2, 4.8 Hz, 1H), 4.41-4.22 (m, 4H), 3.16 (s, 3H), 2.95-2.86 (m, 2H), 2.61-2.33 (m, 5H), 2.08-1.98 (m, 3H), 1.72-1.64 (m, 4H), 1.45-1.29 (m, 7H), 1.27-1.14 (m, 4H).

Example 105: 3-(5-(((1S,2S)-2-(3-(cyclopropylmethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-51)

(Bromomethyl)cyclopropane (105-1, 123 μL, 1.27 mmol) was added to a solution of 3-(5-(((1S,2S)-2-(3-hydroxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (99-2, 70 mg, 0.10 mmol) and AgOTf (163 mg, 0.63 mmol) (mixed with Celite® 1:1) in DCM (1 mL) and the resulting mixture was stirred at rt for 3 h. The reaction mixture was then diluted with DCM, filtered, and concentrated to dryness. The crude material was taken up in 1M HCl (aq) and washed with EtOAc (×3). The acidic layer was basified using NaHCO₃ and extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N) in DCM (with 0.1% Et₃N) to afford I-51 (13 mg, 0.026 mmol, 26% yield) as a pale yellow solid. MS [M+H]⁺=468.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.16 (s, 1H), 7.02 (d, J=8.4 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.38 (dd, J=17.2, 5.5 Hz, 1H), 4.25 (dd, J=17.3, 5.0 Hz, 2H), 3.98-3.84 (m, 1H), 3.51-3.38 (s, 2H), 3.11 (d, J=6.8 Hz, 1H), 3.04-2.77 (m, 2H), 2.59 (d, J=16.6 Hz, 1H), 2.43-2.34 (m, 2H), 2.12-2.02 (m, 1H), 2.02-1.89 (m, 2H), 1.86-1.70 (m, 2H), 1.67-1.50 (m, 2H), 1.48-1.00 (m, 6H), 0.49-0.38 (m, 1H), 0.15-0.08 (m, 1H).

Example 106: 3-(1-oxo-5-(((1S,2S)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-49)

Step 1. Dimethyl 2-diazomalonate (106-3)

To a solution of 4-acetamidobenzenesulfonyl azide (106-2, 6.47 g, 26.1 mmol), TEA (5.8 mL, 42 mmol) and MeCN (80 mL) was added dimethyl malonate (106-1, 2.0 mL, 17 mmol) dropwise at 0° C. and the resulting mixture was stirred vigorously overnight. The reaction mixture was then diluted with Et₂O (200 mL) and filtered. The filtrate was concentrated, the crude material was dissolved in minimal amount of DCM, and excess heptane was added until precipitation of a white solid occurred. The solids were filtered off and the filtrate was concentrated. This sequence was repeated three more times to afford 106-3 (1.92 g, 12.1 mmol, 70% yield) as a pale yellow oil. ¹H NMR (400 MHz, Chloroform-d) δ 3.86 (s, 6H).

Step 2. Dimethyl 2-(2,2,2-trifluoroethoxy)malonate (106-5)

To a solution of dimethyl 2-diazomalonate (106-3, 500 mg, 3.16 mmol), TFE (106-4, 1.04 mL, 15.8 mmol) and DCM (4 mL) under an atmosphere of nitrogen was added rhodium (II) acetate (12.5 mg, 0.032 mmol) in a single portion and the resulting mixture was stirred at rt overnight. The reaction mixture was concentrated to dryness and the crude material purified via silica gel chromatography eluting with 100% DCM to afford 106-5 (640 mg, 2.80 mmol, 88% yield) as a yellow oil. ¹H NMR (400 MHz, Chloroform-d) δ 4.73 (s, 1H), 4.11 (q, J=8.5 Hz, 2H), 3.84 (s, 6H).

Step 3. 2-(2,2,2-trifluoroethoxy)propane-1,3-diol (106-6)

To a solution of dimethyl 2-(2,2,2-trifluoroethoxy)malonate (106-5, 640 mg, 2.8 mmol) in THF (10 mL) under an atmosphere of nitrogen was added 2M LiAlH₄ in THF (2.8 mL, 5.7 mmol) dropwise at 0° C. The resulting mixture was stirred at 30 min at 0° C. and then at rt overnight. The reaction mixture was cooled to 0° C., diluted with THF (20 mL), and Na₂SO₄.10H₂O (900 mg) was added slowly portion wise. After stirring for 10 min, Na₂SO₄ (approx. 300 mg) was added stirring was continued vigorously for 10 min and then the rest of Na₂SO₄.10H₂O (444 mg) was added portion wise. Additional Na₂SO₄ (approx. 300 mg) was added and stirring was continued for 3 hours at rt. The reaction mixture was filtered through a pad of Celite® (washed with minimal amount of EtOAc). The filtrate was concentrated to dryness to afford 106-6 (273 mg, 1.57 mmol, 56% yield) as a colorless oil, which was carried onto the next step without purification. ¹H NMR (400 MHz, DCM-d₂) δ 4.04 (q, J=8.8 Hz, 2H), 3.77-3.66 (m, 4H), 3.65-3.58 (m, 1H).

Step 4. 2-(2,2,2-trifluoroethoxy)propane-1,3-diyl dimethanesulfonate (106-7)

To a solution of 2-(2,2,2-trifluoroethoxy)propane-1,3-diol (106-6, 273 mg, 1.57 mmol), DIPEA (0.68 mL, 3.9 mmol), DMAP (19 mg, 0.16 mmol) in DCM (5 mL) was added MsCl (0.27 mL, 3.5 mmol) dropwise at 0° C. and the resulting mixture was stirred at rt for 4 h. The reaction mixture was then quenched with saturated aqueous NaHCO₃ at 0° C. and extracted with DCM (×2). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to afford 106-7. The crude material was taken onto the next step without purification.

Step 5. 3-(1-oxo-5-(((1S,2S)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-49)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 380 mg, 1.06 mmol) and DIPEA (1.1 mL, 6.3 mmol) in MeCN (1 mL) was added a solution of 2-(2,2,2-trifluoroethoxy)propane-1,3-diyl dimethanesulfonate (106-7, 527 mg, 1.60 mmol) in MeCN (1 mL). The resulting mixture was stirred at 140° C. for 14 h in the μW and then concentrated. The crude material was purified via silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM to afford I-49 (41.6 mg, 0.081 mmol, 8% yield) as a white powder. MS [M+H]⁺=496.2. ¹H NMR (400 MHz, DCM-d₂) δ 8.75 (s, 1H), 7.70 (dd, J=8.7, 4.4 Hz, 1H), 7.03-6.86 (m, 2H), 5.11 (ddd, J=13.5, 8.5, 5.2 Hz, 1H), 4.39-4.18 (m, 3H), 4.13 (quint, J=5.9 Hz, 1H), 3.75 (q, J=8.8 Hz, 2H), 3.62 (dq, J=14.7, 6.6 Hz, 2H), 3.24 (q, J=7.2 Hz, 1H), 3.04 (t, J=6.7 Hz, 1H), 2.91-2.74 (m, 2H), 2.49-2.38 (m, 1H), 2.37-2.25 (m, 1H), 2.22-2.13 (m, 1H), 2.09-2.01 (m, 1H), 1.92-1.83 (m, 1H), 1.79-1.66 (m, 2H), 1.45-1.10 (m, 4H).

Example 107: 3-(5-(((1S,2S)-2-(3-(2,2-difluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-50)

Step 1. Dimethyl 2-(2,2-difluoroethoxy)malonate (107-2)

To dimethyl 2-diazomalonate (106-3, 500 mg, 3.16 mmol), 2,2-difluoroethan-1-ol (107-1, 1297 mg, 15.8 mmol) and DCM (4 mL) under nitrogen atmosphere was added rhodium (II) acetate (12.5 mg, 0.032 mmol) in a single portion. The reaction mixture was stirred at rt overnight. The reaction mixture was concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 100% DCM to afford 107-2 (470 mg, 2.22 mmol, 70% yield) as a yellow oil. ¹H NMR (400 MHz, Chloroform-d) δ 6.00 (tt, J=55.2, 4.2 Hz, 1H), 4.76 (t, J=6.4 Hz, 1H), 3.96-3.87 (m, 2H), 3.85 (s, 6H).

Step 2. 2-(2,2-difluoroethoxy)propane-1,3-diol (107-3)

To dimethyl 2-(2,2-difluoroethoxy)malonate (107-2, 470 mg, 2.22 mmol) in THF (5 mL) under nitrogen atmosphere was added 2M LiAlH₄ in THF (2.2 mL, 4.4 mmol) dropwise at 0° C. The reaction mixture was stirred for 30 min at 0° C. and at rt overnight. The reaction mixture was cooled to 0° C., diluted with THF (20 mL) and Na₂SO₄.10H₂O (1071 mg) was added slowly portionwise. The reaction mixture was stirred vigorously for 1 hour. Celite R was added to the reaction mixture until the solids were free flowing and the reaction mixture was stirred for 40 min at rt. Suspension was filtered, washing with THF (minimal) and concentrated to dryness to afford 107-3 (147 mg, 0.942 mmol, 43% yield) as a colorless oil. ¹H NMR (400 MHz, DCM-d₂) δ 6.17-5.78 (m, 1H), 3.97-3.82 (m, 2H), 3.81-3.57 (m, 5H).

Step 3. 2-(2,2-difluoroethoxy)propane-1,3-diyl dimethanesulfonate (107-4)

To 2-(2,2-difluoroethoxy)propane-1,3-diol (107-3, 147 mg, 0.942 mmol), DIPEA (0.41 mL, 2.4 mmol), and DMAP (11.5 mg, 0.094 mmol) in DCM (5 mL) was added MsCl (0.16 mL, 2.07 mmol) dropwise at 0° C. The resulting mixture was stirred at rt for 4 h. The reaction mixture was then quenched with saturated aqueous NaHCO₃ at 0° C. and then extracted with DCM (×2). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness to afford 107-4, which was taken onto the next step without purification.

Step 4. 3-(5-(((1S,2S)-2-(3-(2,2-difluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-50)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 220 mg, 0.62 mmol), DIPEA (0.64 mL, 3.7 mmol) and MeCN (1 mL) was added a solution of 2-(2,2-difluoroethoxy)propane-1,3-diyl dimethanesulfonate (107-4, 288 mg, 0.92 mmol) in MeCN (1 mL). The resulting mixture was stirred for 14 hours at 140° C. in the μW. The reaction mixture was then concentrated and the crude material was purified via silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM to afford I-50 (20 mg, 0.04 mmol, 7% yield) as a white solid. MS [M+H]⁺=478.5. ¹H NMR (400 MHz, DCM-d₂) δ 8.31 (s, 1H), 7.86-7.46 (m, 1H), 7.08-6.81 (m, 2H), 5.81 (tt, J=55.3, 3.9 Hz, 1H), 5.22-5.01 (m, 1H), 4.39-4.14 (m, 3H), 4.06 (quint, J=5.8 Hz, 1H), 3.68-3.45 (m, 4H), 3.33-3.09 (m, 1H), 3.06-2.93 (m, 1H), 2.89-2.72 (m, 2H), 2.46-2.35 (m, 1H), 2.35-2.24 (m, 1H), 2.21-2.11 (m, 1H), 2.07-1.99 (m, 1H), 1.92-1.81 (m, 1H), 1.76-1.65 (m, 2H), 1.45-1.22 (m, 3H), 1.19-1.07 (m, 1H).

Example 108: 3-(5-(((1S,2S)-2-(3-(3,3-difluorocyclobutoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-56)

Step 1. Dimethyl 2-(3,3-difluorocyclobutoxy)malonate (108-2)

To dimethyl 2-diazomalonate (106-3, 500 mg, 3.16 mmol), 3,3-difluorocyclobutan-1-ol (108-1, 513 mg, 4.74 mmol), and DCM (4 mL) under an atmosphere of nitrogen was added rhodium (II) acetate (12.5 mg, 0.032 mmol) in a single portion. The reaction mixture was stirred at rt overnight and then concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 100% DCM to afford 108-2 (649 mg, 2.72 mmol, 86% yield) as a yellow oil. ¹H NMR (400 MHz, Chloroform-d) δ 4.47 (s, 1H), 4.19-4.12 (m, 1H), 3.82 (s, 6H), 2.95-2.84 (m, 2H), 2.77-2.66 (m, 2H).

Step 2. 2-(3,3-difluorocyclobutoxy)propane-1,3-diol (108-3)

To dimethyl 2-(3,3-difluorocyclobutoxy)malonate (108-2, 744 mg, 3.10 mmol) in THF (6 mL) under an atmosphere of nitrogen was added 2M LiAlH₄ in THF (3.1 mL, 6.3 mmol) dropwise at 0° C. The resulting mixture was stirred for 30 min at 0° C. and then at rt overnight. The reaction mixture was cooled to 0° C., diluted with THF (20 mL), and Na₂SO₄.10H₂O (1071 mg) was added slowly portionwise. The resulting mixture was stirred vigorously for 1 hour and Celite® was added until the solids were free flowing. The reaction mixture was stirred for 40 min at rt. The suspension was filtered, washed with THF (minimal), and the filtrate was concentrated to dryness to afford 108-3 (499 mg, 2.74 mmol, 88% yield) as a colorless oil. ¹H NMR (400 MHz, DCM-d₂) δ 4.35-4.12 (m, 1H), 3.86-3.75 (m, 1H), 3.71-3.59 (m, 4H), 3.48-3.39 (m, 1H), 2.91-2.81 (m, 2H), 2.60-2.46 (m, 2H), 2.26-2.11 (m, 1H).

Step 3. 2-(3,3-difluorocyclobutoxy)propane-1,3-diyl dimethanesulfonate (108-4)

To 2-(3,3-difluorocyclobutoxy)propane-1,3-diol (108-3, 483 mg, 2.65 mmol), DIPEA (1.2 mL, 6.6 mmol), and DMAP (32 mg, 0.27 mmol) in DCM (5 mL) was added MsCl (0.45 mL, 5.8 mmol) dropwise at 0° C. The resulting mixture was stirred at rt for 4 h and then quenched with saturated aqueous NaHCO₃ at 0° C. and extracted with DCM (×2). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness to afford 108-4, which was taken onto the next step without purification.

Step 4. 3-(5-(((1S,2S)-2-(3-(3,3-difluorocyclobutoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-56)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 450 mg, 1.26 mmol), DIPEA (1.3 mL, 7.5 mmol), and MeCN (2.5 mL) was added a solution of 2-(3,3-difluorocyclobutoxy)propane-1,3-diyl dimethanesulfonate (108-4, 852 mg, 2.52 mmol) in MeCN (1 mL). The resulting mixture was stirred for 14 hours at 140° C. in the μW and then concentrated. The crude material was purified via silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM to afford I-56 (30 mg, 0.058 mmol, 5% yield) as a white solid. MS [M+H]⁺=504.4. ¹H NMR (400 MHz, DCM-d₂) δ 8.79 (s, 1H), 7.71 (s, 1H), 7.18-6.69 (m, 2H), 5.11 (ddd, J=13.5, 8.6, 5.1 Hz, 1H), 4.39-4.16 (m, 3H), 4.05-3.93 (m, 1H), 3.92-3.81 (m, 1H), 3.61 (s, 2H), 3.29-3.13 (m, 1H), 3.07-2.91 (m, 1H), 2.91-2.72 (m, 4H), 2.57-2.39 (m, 3H), 2.39-2.24 (m, 1H), 2.21-2.13 (m, 1H), 2.10-2.00 (m, 1H), 1.92-1.83 (m, 1H), 1.77-1.68 (m, 2H), 1.48-1.12 (m, 4H).

Example 109: 4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile HC(O)OH Salt (I-129) and 4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)(methyl)amino)methyl)benzonitrile HC(O)OH Salt (I-130)

Step 1. 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)-1-(hydroxymethyl)piperidine-2,6-dione (109-2)

To a stirred solution of tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)carbamate (109-1, 860 mg, 1.46 mmol) in THF (4 mL) was added 4M hydrogen chloride in dioxane (8.0 mL, 32 mmol) and the resulting mixture stirred for 8 hours at 60° C. The reaction mixture was filtered and the obtained solid was washed with Et₂O (×3) and vacuum dried overnight to afford crude hydrochloride salt of 109-2 (525 mg, 1.08 mmol, 74% yield) as a white solid. The product was used in the next step without further purification. MS [M+2H]⁺=389.3.

Step 2. 4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile HC(O)OH Salt (I-129) and 4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)(methyl)amino)methyl)benzonitrile HC(O)OH Salt (I-130)

To a stirred solution of crude 109-2 (180 mg, 0.425 mmol), NaBH(OAc)₃ (135 mg, 0.637 mmol), and DMF (1 mL) under an atmosphere of nitrogen was added 4-formylbenzonitrile (109-3, 58.5 mg, 0.446 mmol) in one portion and the resulting mixture was stirred 2 h at room temperature and then concentrated. The crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collected fractions contained ˜2 drops of formic acid) to afford the formate salt of I-129 (31 mg, 0.060 mmol, 14% yield) as a white solid and the formate salt of I-130 (25 mg, 0.048 mmol, 11% yield) as a white solid. Data for I-129: MS [M+H]⁺=473.1. ¹H NMR (400 MHz, MeCN-d₃) δ 8.88 (br s, 1H), 8.10 (s, 1H), 7.71-7.58 (m, 3H), 7.50-7.41 (m, 2H), 7.13-7.07 (m, 1H), 7.03 (dd, J=8.4, 2.3 Hz, 1H), 5.04 (ddd, J=13.4, 5.2, 2.1 Hz, 1H), 4.37-4.18 (m, 3H), 3.94 (d, J=14.8 Hz, 1H), 3.87 (d, J=14.7 Hz, 1H), 2.88-2.67 (m, 3H), 2.18-1.98 (m, 3H), 1.77-1.66 (m, 2H), 1.48-1.16 (m, 4H). Data for I-130: MS [M+H]⁺=487.1. ¹H NMR (400 MHz, DCM-d₂) δ 8.28 (s, 1H), 7.73 (d, J=8.4 Hz, 1H), 7.53 (dd, J=8.3, 2.5 Hz, 2H), 7.39 (t, J=8.3 Hz, 2H), 7.08-6.92 (m, 2H), 5.19-5.05 (m, 1H), 4.42-4.23 (m, 3H), 3.86 (d, J=14.6 Hz, 1H), 3.77 (dd, J=14.6, 3.8 Hz, 1H), 2.97-2.72 (m, 3H), 2.41-2.25 (m, 1H), 2.22 (d, J=2.3 Hz, 3H), 2.25-2.14 (m, 2H), 2.02-1.91 (m, 1H), 1.83-1.67 (m, 2H), 1.53-1.20 (m, 4H).

Example 110: 3-(5-(((1S,2S)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-205)

Step 1. 2-isopropoxypropane-1,3-diol & 3-isopropoxypropane-1,2-diol (183-2 and 183-3)

To a solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methanol (183-1, 3.00 g, 22.7 mmol) in DCM (100 mL) was added triethyl silane (4.39 g, 27.2 mmol) and ethyl aluminum dichloride (25% in toluene (25.4 mL, 49.9 mmol) at −30° C. and the resulting mixture was stirred at rt for 2 h. The reaction mixture was quenched with aq. NaHCO₃ solution and extracted with DCM (3×100 mL). The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness to afford a mixture of crude 183-2 and 183-3 (1.3 g, crude) as colorless liquid. The crude material was taken onto the next step without purification.

Step 2. 2-isopropoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) & 3-isopropoxypropane-1,2-diyl bis(4-methylbenzenesulfonate) (183-4 and 183-5)

To a solution of a mixture of 183-2 and 183-3 (1.3 g, 9.6 mmol) in CH₃CN (30 mL) was added Et₃N (5.3 mL, 38 mmol), DMAP (0.04 g, 0.3 mmol) and TsCl (4.00 g, 21.1 mmol) at 0° C. and resulting mixture was stirred at room temperature for 16 h. The reaction mixture was then filtered and the filtrate was concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 10% EtOAc in hexanes to afford a mixture of 183-4 and 183-5 (3.1 g, 7.0 mmol, 73% yield) as a pale brown liquid. MS [M+H+18]⁺=460.1.

Step 3. 3-(5-(((1S,2S)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-205)

To a solution of I-12 (400 mg, 1.2 mmol) in MeCN (10 mL) was added a mixture of 183-4 and 183-5 (670 mg, 1.5 mmol) and DIPEA (0.62 mL, 3.5 mmol) and resulting mixture was stirred at 120° C. for 16 h under microwave irradiation. The reaction mixture was then diluted with sat. sodium bicarbonate solution and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified via preparative TLC in 3% MeOH in DCM as eluent to afford compound I-205 (20 mg, 0.05 mmol, 4% yield) as an off-white solid. MS [M+H]⁺=442.4. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.15 (s, 1H), 7.03 (d, J=8.4 Hz, 1H), 5.08 (dd, J=13.2, 4.8 Hz, 1H), 4.80-4.69 (m, 1H), 4.40-4.22 (m, 2H), 3.61-3.49 (m, 1H), 3.15-2.85 (m, 2H), 2.60-2.31 (m, 3H), 2.25-1.98 (m, 3H), 1.95-1.66 (m, 5H), 1.50-1.49 (m, 2H), 1.07-1.01 (m, 7H).

Example 111: 3-(5-(((1S,2S)-2-(ethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-79) HC(O)OH Salt

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 103 mg, 0.25 mmol), acetaldehyde (2-1, 0.021 mL, 0.37 mmol) and trifluoroethanol (1 mL) under an atmosphere of nitrogen was added NaBH(OAc)₃ (78 mg, 0.37 mmol) in one portion. The resulting mixture was stirred at rt for 1 h and then diluted with DCM and concentrated onto Celite®. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% Et₃N) in DCM and then further purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collected fractions contained ˜3 drops of formic acid). The fractions containing the desired product were combined and lyophilized to afford the formate salt of I-79 (30 mg, 0.067 mmol, 30% yield). MS [M+H]⁺=386.1. ¹H NMR (400 MHz, DMSO-d₆) δ 8.29 (s, 2H), 7.61 (d, J=8.3 Hz, 1H), 7.21 (d, J=1.9 Hz, 1H), 7.08 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.43-4.33 (m, 1H), 4.30-4.19 (m, 2H), 2.97-2.84 (m, 1H), 2.70 (ddd, J=18.2, 7.7, 3.3 Hz, 2H), 2.63-2.55 (m, 2H), 2.42-2.26 (m, 1H), 2.08-2.04 (m, 1H), 2.02-1.90 (m, 2H), 1.71-1.61 (m, 2H), 1.41-1.15 (m, 4H), 1.00 (t, J=7.1 Hz, 3H).

Example 112: 3-(5-(((1S,2S)-2-(isopropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-131)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 90 mg, 0.25 mmol), acetone (0.09 mL, 1 mmol), and MgSO₄ (61 mg, 0.50 mmol) in DMF (1.0 mL) was added NaBH(OAc)₃ (107 mg, 0.50 mmol) in one portion and the resulting mixture was stirred overnight at room temperature. The reaction mixture was then filtered and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0 to 100% EtOH:EtOAc (v/v=3:1, with 1% Et₃N) in DCM to afford I-131 (95 mg, 0.24 mmol, 94% yield) as a white solid. MS [M+H]⁺=400.1. ¹H NMR (400 MHz, DCM-d₂) δ 7.67 (d, J=8.3 Hz, 1H), 7.07-6.87 (m, 2H), 5.30 (br s, 1H), 5.04 (dd, J=13.3, 5.2 Hz, 1H), 4.29-4.21 (m, 1H), 4.15 (tt, J=9.3, 4.6 Hz, 1H), 2.98 (heptd, J=6.3, 1.8 Hz, 1H), 2.87-2.80 (m, 1H), 2.78-2.67 (m, 2H), 2.30-2.14 (m, 1H), 2.13-1.99 (m, 3H), 1.91 (s, 1H), 1.76-1.62 (m, 2H), 1.41-1.16 (m, 4H), 1.11-0.99 (m, 6H).

Example 113: 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile (I-132)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 30 mg, 0.074 mmol) in DMF (0.5 mL) was added 3-formylbenzonitrile (113-1, 10 mg, 0.076) and sodium triacetoxyborohydride (32 mg, 0.15 mmol) and the resulting mixture was stirred at rt for 2 h. Additional 3-formylbenzonitrile (113-1, 5 mg) was added and stirring was continued at rt for 4 h. Additional sodium triacetoxyborohydride (40 mg) was added and the reaction mixture was stirred at rt for 16 h. Further sodium triacetoxyborohydride (30 mg) was added and stirring was continued at 60° C. for 2 h. The crude reaction mixture was diluted with DMSO (1 mL) and purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-132 (9.0 mg, 0.017 mmol, 23% yield). MS [M+H]⁺=473.3. ¹H NMR (400 MHz, MeCN-d₃) δ 8.83 (s, 1H), 7.68 (d, J=1.7 Hz, 1H), 7.65-7.53 (m, 3H), 7.44 (t, J=7.7 Hz, 1H), 7.11-7.08 (m, 1H), 7.06-7.01 (m, 1H), 5.04 (ddd, J=13.4, 5.2, 2.7 Hz, 1H), 4.40-4.27 (m, 2H), 4.26-4.15 (m, 1H), 3.90 (d, J=14.5 Hz, 1H), 3.83 (d, J=14.5 Hz, 1H), 2.88-2.64 (m, 3H), 2.40 (qd, J=13.2, 4.9 Hz, 1H), 2.07-2.01 (m, 1H), 1.81-1.65 (m, 2H), 1.47-1.17 (m, 4H).

Example 114: 3-(5-(((1S,2S)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-133)

To 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 80 mg, 0.22 mmol) and 3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (51-3, 38.3 mg, 0.34 mmol) in TFE (1.0 mL) was added NaBH(OAc)₃ (316 mg, 0.45 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h, then filtered, and concentrated. The crude material was purified via silica gel chromatography eluting with 0 to 100% EtOH:EtOAc (v/v=1:3, with 1% NEt₃) in DCM to afford I-133 (72 mg, 0.16 mmol, 70% yield) as a white solid. MS [M+H]⁺=456.2. ¹H NMR (400 MHz, DCM-d₂) δ 7.68 (d, J=8.2 Hz, 1H), 7.02-6.96 (m, 2H), 5.07 (dd, J=13.3, 5.2 Hz, 1H), 4.37-4.20 (m, 2H), 4.18-4.06 (m, 1H), 2.97 (d, J=12.6 Hz, 1H), 2.83 (d, J=12.7 Hz, 1H), 2.79-2.59 (m, 3H), 2.36-2.17 (m, 1H), 2.17-2.06 (m, 2H), 2.05-1.96 (m, 1H), 1.89 (d, J=2.7, Hz, 6H), 1.75-1.65 (m, 2H), 1.36-1.13 (m, 4H).

Example 115: 3-(5-((2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-134)

Step 1. 5-((2-oxocyclohexyl)oxy)isobenzofuran-1(3H)-one (115-3)

5-hydroxyisobenzofuran-1(3H)-one (115-2, 415 mg, 2.76 mmol), 2-bromocyclohexanone (115-1, 508 mg, 2.87 mmol), and potassium carbonate (807 mg, 5.84 mmol) were treated with DMF (10 mL) and the resulting mixture was stirred at rt for 2 days. The reaction mixture was then diluted with ethyl acetate (150 mL) and washed with 0.5 M LiCl (aq) (2×20 mL), saturated aqueous sodium bicarbonate solution (30 mL) and brine (25 mL). The organic phase was collected, dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc in hexane to afford 115-3 (471 mg, 1.72 mmol, 62% yield) as a beige solid. MS [M+H]⁺=247.2. ¹H NMR (400 MHz, DCM-d) δ 7.84 (d, J=8.5 Hz, 1H), 7.06 (d, J=8.5 Hz, 1H), 6.90 (s, 1H), 5.29 (d, J=3.5 Hz, 2H), 4.87 (dd, J=10.7, 5.7 Hz, 1H), 2.64 (d, J=13.3 Hz, 1H), 2.52 (t, J=13.7 Hz, 2H), 2.26-2.00 (m, 3H), 1.87 (q, J=12.5 Hz, 2H).

Step 2. 5-(((1R,2R)-2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)isobenzofuran-1(3H)-one or 5-(((1S,2S)-2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)isobenzofuran-1 (3H)-one (115-5)

5-((2-oxocyclohexyl)oxy)isobenzofuran-1(3H)-one (115-3, 151 mg, 0.613 mmol), 3,3-difluoropyrrolidine hydrochloride (115-4, 101 mg, 0.704 mmol), and sodium triacetoxyborohydride (194 mg, 0.915 mmol) were dissolved in DMF (2 mL) under an atmosphere of nitrogen and the resulting mixture stirred at rt overnight. The reaction mixture was then diluted with ethyl acetate (80 mL) and water (20 mL). The mixture was basified with saturated aqueous sodium bicarbonate solution. The organic layer was washed with 0.5 M lithium chloride solution (20 mL) and brine (20 mL), dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc (with 0.1% NEt₃) in heptane (with 0.1% NEt₃) to afford a mixture of trans products (58 mg, 0.17 mmol, 28% yield) as a colorless oil. The isolated trans products were separated by chiral SFC (Method 2.1×25.0 cm Chiralpak AD-H, CO₂ Co-solvent: MeOH/i-PrOH (1:1); Isocratic method: 10% Co-solvent at 80 g/min; 100 bar, 25° C.). Peak 1 was isolated to afford a single enantiomer 115-5 (20 mg, 0.059 mmol, 10% yield). MS [M+H]⁺=338.5. ¹H NMR (400 MHz, DCM-d₂) δ 7.81 (d, J=8.4 Hz, 1H), 7.08 (d, J=8.5 Hz, 1H), 6.99 (s, 1H), 5.26 (s, 2H), 4.52-4.31 (m, 1H), 3.10 (dq, J=37.3, 12.1 Hz, 2H), 2.99-2.82 (m, 2H), 2.79-2.62 (m, 1H), 2.30-2.09 (m, 3H), 2.09-1.94 (m, 1H), 1.86-1.71 (m, 2H), 1.51-1.30 (m, 4H). Chiral SFC Peak 1: Rt=1.91 min. Absolute stereochemistry not determined.

Step 3. Single Enantiomer of Ethyl 2-(chloromethyl)-4-((˜2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)benzoate (115-6)

To single enantiomer of 5-((˜2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)isobenzofuran-1(3H)-one (115-5, Peak 1, 20 mg, 0.060 mmol) dissolved in EtOH (1 mL) was added thionyl chloride (30 μL, 0.41 mmol) and the resulting mixture was stirred at 70° C. for 16 h. Additional thionyl chloride (50 μL, 0.69 mmol) was added and stirring was continued at 70° C. for 16 h. The reaction mixture was diluted with water (10 mL) and neutralized with saturated aqueous sodium bicarbonate solution. The aqueous mixture was extracted with ethyl acetate (3×15 mL) and the combined organic phases were washed with brine (10 mL), dried over magnesium sulfate, filtered and concentrated to afford 115-6 (20 mg, 43 μmol, 71% yield) as an amber colored gum. MS [M+H]⁺=402.5.

Step 4. Single Enantiomer of 3-(5-((˜2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-134)

A mixture of single enantiomer of ethyl 2-(chloromethyl)-4-((˜2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)benzoate (115-6, 20.4 mg, 0.051 mmol) in DMF (0.5 mL) and DIPEA (40 μL, 0.23 mmol) was purged with nitrogen three times. 3-Aminopiperidine-2,6-dione hydrochloride (I-1c, 14 mg, 0.083 mmol) was then added in one portion and the reaction mixture was purged with nitrogen again and was stirred at 80° C. for 2 hours and then at 110° C. for 27 h. The reaction mixture was then diluted with ethyl acetate (40 mL) and washed with saturated aqueous sodium bicarbonate solution (2×10 mL) and brine (10 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collected fractions contained ˜3 drops of formic acid). The fractions containing the desired product were combined and lyophilized to afford I-134 (6.0 mg, 0.014 mmol, 27% yield) as a white solid. MS [M+H]⁺=448.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.21 (d, J=2.1 Hz, 1H), 7.07 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.58-4.50 (m, 1H), 4.44-4.20 (m, 2H), 3.20-3.06 (m, 1H), 3.06-2.88 (m, 2H), 2.88-2.80 (m, 2H), 2.66-2.55 (m, 2H), 2.38 (dd, J=13.2, 4.5 Hz, 1H), 2.24-2.08 (m, 2H), 2.05-1.93 (m, 2H), 1.93-1.82 (m, 1H), 1.63 (s, 2H), 1.52-1.18 (m, 4H).

Example 116: 3-(1-oxo-5-(((1S,2S)-2-(3-(pyridin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-135) Step 1: 2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilaundecan-6-ol (116-2)

To a solution of propane-1,2,3-triol (116-1, 5.00 g, 54.3 mmol) in THF (120 mL), Imidazole (8.13 g, 119 mmol) and TBS chloride (16.4 g, 109 mmol) were added at 0° C. and stirred at room temperature for 16 h. The reaction mixture was diluted with cold water, extracted with EtOAc (3×100 mL). The combined organic extracts were washed with water, brine, dried over anhydrous Na₂SO₄ and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 15% EtOAc in Hexane to afford 116-2 (14.0 g, 43.7 mmol, 80%) as a colorless oil. ¹H NMR (400 MHz, DMSO-d₆): δ 4.59-4.61 (m, 1H), 3.52-3.32 (m, 5H), 0.86 (s, 18H), 0.03 (s, 12H).

Step 2: 3-((2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilaundecan-6-yl)oxy)pyridine (116-4)

To a stirred solution of 2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilaundecan-6-ol (116-2, 4.05 g, 12.6 mmol), pyridin-3-ol (116-3, 1.00 g, 10.5 mmol) and PPh₃ (3.30 g, 12.6 mmol) in THF (50 mL), at 0° C. was added DIAD (2.55 g, 12.6 mmol) drop wise and the resulting mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with cold water and extracted with EtOAc (3×50 mL). The combined organic phases were washed with water, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 20-40% EtOAc in Hexane to afford 116-4 (1.60 g, 4.02 mmol, 38%) as a colorless oil. ¹H NMR (300 MHz, CDCl₃): δ 8.35-8.31 (m, 1H), 8.21-8.18 (m, 1H), 7.30-7.15 (m, 2H), 4.39-4.35 (m, 1H), 3.87-3.76 (m, 4H), 0.88 (s, 18H), 0.02 (s, 12H).

Step 3: 2-(pyridin-3-yloxy)propane-1,3-diol (116-5)

To a stirred solution of 3-((2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disilaundecan-6-yl)oxy)pyridine (116-4, 1.60 g, 4.02 mmol) in MeOH (30 mL) at 0° C. was added concentrated HCl (3.20 mL) drop wise and the resulting mixture was stirred at room temperature for 4 h. The reaction mixture was concentrated to dryness. The obtained crude material was diluted with cold water, basified with NaHCO₃, and the aqueous mixture was concentrated to dryness. The residue obtained was stirred with 10% MeOH in DCM, filtered, and the filtrate was concentrated to dryness to afford 116-5 (0.800 g, crude) as pale brown oil. The product was carried on to the next step without purification. [M+H]⁺=170.20.

Step 4: 2-(pyridin-3-yloxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (116-6)

To a solution of 2-(pyridin-3-yloxy)propane-1,3-diol (116-5, 0.80 g, 4.73 mmol) in DCM (40 mL), Et₃N (1.91 g, 18.9 mmol) at 0° C. was added DMAP (0.190 g, 1.56 mmol) and tosyl chloride (2.25 g, 11.8 mmol) and the resulting mixture was stirred at room temperature for 3 h. The reaction mixture was quenched with cold water and extracted with DCM (3×30 mL). The combined organic phases were washed water, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 50% EtOAc in hexane to afford 116-6 (1.30 g, 2.72 mmol, 58%) as colorless oil. MS [M+H]⁺=478.10.

Step 5. 3-(1-oxo-5-(((1S,2S)-2-(3-(pyridin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-135)

To a solution of 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-15, 200 mg, 0.56 mmol) in acetonitrile (10 mL) was added 2-(pyridin-3-yloxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (116-6, 401 mg, 0.84 mmol) and DIPEA (0.43 g, 3.36 mmol) and the resulting mixture was stirred at 120° C. for 16 hr under microwave irradiation. The reaction mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by reverse phase HPLC (Column: LUNA C18 (250 mm×21.2 mm), 5.0 g, Mobile phase-A: 0.01% HCOOH (aq), Mobile phase-B: acetonitrile; Method: 0/10, 2/10, 8/50. Flow rate: 20 mL/min). The fractions containing the desired product were combined and lyophilized to afford I-135 (25 mg, 0.05 mmol, 9%) as white solid. MS [M+H]⁺=491.20. ¹H NMR (400 MHz, DMSO-d₆): δ 10.96 (s, 1H), 8.18-8.13 (m, 2H), 7.62 (d, J=8.4 Hz, 1H), 7.31-7.23 (m, 2H), 7.17 (s, 1H), 7.05 (d, J=8.4 Hz, 1H), 5.09 (dd, J=13.2, 4.8 Hz, 1H), 4.81-4.78 (m, 1H), 4.41-4.22 (m, 3H), 3.78-3.73 (m, 2H), 3.24-3.21 (m, 2H), 3.09-3.08 (m, 1H), 2.94-2.87 (m, 1H), 2.85-2.32 (m, 4H), 2.07-1.98 (m, 2H), 1.98-1.81 (m, 1H), 1.66-1.64 (m, 2H), 1.36-1.23 (m, 2H).

Example 117: 3-(5-(((1S,2S)-2-(ethyl((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-136)

To a solution of 3-(5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-29, 12 mg, 0.032 mmol) in DCM (0.5 mL) was added 3-fluorobicyclo[1.1.1]pentane-1-carbaldehyde (51-3, 7.4 mg, 0.065 mmol). Sodium triacetoxyborohydride (21 mg, 0.097 mmol) was added and the resulting mixture was stirred at rt for 1 h. The reaction mixture was then concentrated to dryness and the crude material was purified by reverse phase HPLC (Method Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 35-60% MeCN, 3.5 min gradient; collected fractions contained several drops of formic acid). The fractions containing the desired product were combined and lyophilized to afford I-136 (6.0 mg, 11 μmol, 34% yield) as a white solid. MS [M+H]⁺=470.2. ¹H NMR (400 MHz, DCM-d₂) δ 8.01 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.01 (t, J=2.4 Hz, 1H), 6.97 (dd, J=8.4, 2.2 Hz, 1H), 5.04 (ddd, J=13.3, 5.1, 2.2 Hz, 1H), 4.60-4.48 (s, 1H), 4.32-4.17 (m, 2H), 3.35-3.23 (m, 1H), 2.84-2.67 (m, 4H), 2.61-2.50 (m, 2H), 2.34-2.19 (m, 1H), 2.15-2.04 (m, 1H), 1.97-1.83 (m, 8H), 1.75-1.59 (m, 3H), 1.55-1.41 (m, 1H), 0.94 (t, J=7.1 Hz, 3H).

Example 118: 3-(1-oxo-5-(((1S,2S)-2-(4-oxopiperidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-137)

3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 99 mg, 0.14 mmol) was dissolved in DMA (1 mL) and DIPEA (0.07 mL, 0.4 mmol) was added, followed by a solution of 1,5-dichloropentan-3-one (78-1, 21 mg, 0.14 mmol) in DMA (1 mL). The resulting solution was stirred at 85° C. for 2 h and then at rt overnight. Additional 1,5-dichloropentan-3-one (78-1, 21 mg, 0.14 mmol) in DMA (200 uL) was added and stirring was continues at 85° C. for 2 h. The reaction mixture was then cooled to rt, DIPEA (0.07 mL, 0.4 mmol) was added and the mixture was heated to 85° C. for 5 h. The reaction mixture was poured into saturated aqueous NaHCO₃ and extracted with DCM: i-PrOH (4:1). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated. The crude material was purified via silica gel chromatography eluting with 0-10% i-PrOH in DCM to afford I-137 (11 mg, 0.024 mmol, 18% yield) as an off-white solid. MS [M+H]⁺=426.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.23-7.18 (m, 1H), 7.08-7.03 (m, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.83-4.76 (m, 1H), 4.43-4.33 (m, 1H), 4.33-4.22 (m, 1H), 3.16-3.07 (m, 1H), 2.97-2.85 (m, 1H), 2.78 (t, J=6.1 Hz, 4H), 2.63-2.55 (m, 1H), 2.44-2.29 (m, 5H), 2.18-2.04 (m, 1H), 2.03-1.92 (m, 2H), 1.77-1.61 (m, 3H), 1.61-1.49 (m, 1H).

Example 119: 3-(5-(((1S,2S)-2-(4-hydroxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-138)

3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 70.0 mg, 0.153 mmol) was dissolved in MeCN (1 mL) and DIPEA (0.16 mL, 0.92 mmol) was added, followed by a solution of 3-hydroxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (44-2, 203 mg, 0.459 mmol) in MeCN (1 mL) and the resulting solution was stirred at 75° C. overnight. The reaction mixture was then concentrated and the crude material was purified by reverse phase HPLC (Method Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collected fractions contained ˜3 drops of formic acid) to afford the formate salt of I-138 (11.0 mg, 22.0 μmol, 14% yield) as a white solid. MS [M+H]⁺=442.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.17 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.1 Hz, 1H), 7.03 (dd, J=8.4, 1.6 Hz, 1H), 5.10-5.02 (m, 1H), 4.73-4.66 (m, 1H), 4.38 (dd, J=17.1, 9.3 Hz, 1H), 4.25 (dd, J=17.2, 7.9 Hz, 1H), 4.06 (s, 1H), 2.97-2.84 (m, 2H), 2.64-2.55 (m, 1H), 2.48-2.34 (m, 5H), 2.11-1.87 (m, 3H), 1.72-1.55 (m, 3H), 1.54-1.37 (m, 5H), 1.07 (s, 3H).

Example 120: 3-(5-(((1S,2S)-2-(3-hydroxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-139)

To 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 56.0 mg, 0.163 mmol) dissolved in MeCN (1 mL) was added DIPEA (0.17 mL, 0.98 mmol) followed by a solution of 1,4-dichlorobutan-2-ol (139-1, 70.0 mg, 0.489 mmol) in MeCN (1 mL) and the resulting mixture was stirred at 75° C. overnight. The reaction mixture was then cooled to rt and additional DIPEA (0.17 mL, 0.98 mmol) and 1,4-dichlorobutan-2-ol (139-1, 70.0 mg, 0.489 mmol) were added. The reaction mixture was stirred at 75° C. overnight and then concentrated. The crude material was purified by reverse phase HPLC (Method Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient; collected fractions contained ˜3 drops of formic acid) to afford the formate salt of I-139 (13 mg, 25 μmol, 16% yield) as a white solid. MS [M+H]⁺=414.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.20 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.17-7.13 (m, 1H), 7.02 (dd, J=8.2, 2.2 Hz, 1H), 5.10-5.02 (m, 1H), 4.73-4.59 (m, 2H), 4.39 (dd, J=17.2, 8.1 Hz, 1H), 4.25 (dd, J=17.1, 6.7 Hz, 1H), 4.18-4.10 (m, 1H), 2.96-2.85 (m, 1H), 2.80-2.71 (m, 2H), 2.65-2.54 (m, 2H), 2.46-2.35 (m, 3H), 2.19-2.08 (m, 1H), 2.02-1.85 (m, 3H), 1.73-1.45 (m, 5H).

Example 120: 3-(5-(((1S,2S)-2-(3-(benzyloxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-52)

To 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 65.0 mg, 0.142 mmol) dissolved in MeCN (1 mL) was added DIPEA (0.10 mL, 0.57 mmol), followed by a solution of (((1,3-dibromopropan-2-yl)oxy)methyl)benzene (120-1, 109 mg, 0.354 mmol) in MeCN (1 mL). The resulting solution was stirred at 75° C. Upon complete consumption of starting materials, the reaction mixture was cooled to rt, azeotroped with toluene (2×), and concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-52 (4.70 mg, 8.60 μmol, 6% yield) as a glassy solid. MS [M+H]⁺=490.6. ¹H NMR (400 MHz, Chloroform-d) δ 8.11-8.03 (m, 1H), 7.81-7.75 (m, 1H), 7.40-7.32 (m, 3H), 7.32-7.27 (m, 2H), 6.98-6.92 (m, 2H), 5.25-5.14 (m, 1H), 5.06-4.99 (m, 1H), 4.55-4.40 (m, 5H), 4.40-4.26 (m, 2H), 3.83 (s, 1H), 3.73-3.59 (m, 2H), 2.97-2.77 (m, 2H), 2.43-2.27 (m, 2H), 2.26-2.18 (m, 1H), 2.18-2.08 (m, 1H), 1.99-1.87 (m, 1H), 1.87-1.68 (m, 3H).

Example 121: 3-(5-(((1S,2S)-2-(3-hydroxy-3-methylazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HCl Salt (I-141)

2-(chloromethyl)-2-methyloxirane (121-1, 20.0 mg, 0.188 mmol) was added to a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 56.0 mg, 0.163 mmol) in TFE (0.75 mL). The resulting mixture was stirred at rt over 3 days, then heated to 50° C., and stirred at 50° C. for 6 h. The reaction mixture was then heated to 70° C. overnight with stirring, cooled to rt and concentrated to dryness. The crude material was triturated with acetone (×3) and the resultant solid dried under vacuum to afford the HCl salt of I-141 (28.0 mg, 57.0 μmol, 35% yield) as an orange solid. MS [M+H]⁺=414.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.67 (d, J=8.4 Hz, 1H), 7.20 (s, 1H), 7.09 (d, J=8.3 Hz, 1H), 5.14-5.01 (m, 1H), 4.94-4.75 (m, 1H), 4.47-4.36 (m, 1H), 4.34-4.18 (m, 1H), 4.17-3.81 (m, 4H), 2.98-2.83 (m, 1H), 2.64-2.55 (m, 1H), 2.45-2.35 (m, 2H), 2.21-2.10 (m, 1H), 2.03-1.94 (m, 1H), 1.87-1.53 (m, 5H), 1.51-1.30 (m, 3H).

Example 122: 3-(5-(((1S,2S)-2-(isobutylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-142)

To 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 80.0 mg, 0.175 mmol) dissolved in TFE (1.5 mL) was added isobutyraldehyde (17-1a, 16 μL, 0.18 mmol) followed by sodium triacetoxyborohydride (74 mg, 0.35 mmol) and the resulting mixture was stirred at rt overnight. Additional isobutyraldehyde (17-1a, 16 μL, 0.18 mmol), TFE (1 mL), and sodium triacetoxyborohydride (74 mg, 0.35 mmol) were added until complete consumption of I-12 was observed. The reaction mixture was then poured into saturated NaHCO₃ (aq) (2 mL) and extracted with DCM (3×20 mL). The combined organic phases were concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-10% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-142 (45.0 mg, 0.107 mmol, 61% yield) as a white solid. MS [M+H]⁺=400.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.21-7.16 (m, 1H), 7.04 (d, J=8.5 Hz, 1H), 5.07 (dd, J=13.1, 4.7 Hz, 1H), 4.58 (s, 1H), 4.38 (dd, J=17.1, 9.5 Hz, 1H), 4.26 (dd, J=17.1, 8.3 Hz, 1H), 3.10 (s, 1H), 2.97-2.84 (m, 1H), 2.63-2.53 (m, 1H), 2.44-2.34 (m, 2H), 2.18-2.05 (m, 1H), 2.02-1.87 (m, 2H), 1.79-1.54 (m, 5H), 1.53-1.38 (m, 1H), 0.87 (d, J=6.5 Hz, 6H).

Example 123: 3-(5-(((1S,2S)-2-(ethyl(methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-143)

To a solution of 3-(5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-29, 30.0 mg, 81.0 μmol) in DCE (1 mL) was added paraformaldehyde (13.0 mg, 0.433 mmol) followed by sodium triacetoxyborohydride (20.0 mg, 94.0 μmol) and acetic acid (1 μL, 17.0 μmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was then poured into saturated NaHCO₃(aq) (2 mL) and extracted with i-PrOH:DCM (1:9) (×4). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-20% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-143 (16.0 mg, 39.0 μmol, 49% yield) as a white solid. MS [M+H]⁺=386.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.19 (s, 1H), 7.05 (d, J=8.5 Hz, 1H), 5.07 (dd, J=13.2, 5.1 Hz, 1H), 4.69 (s, 1H), 4.39 (dd, J=17.1, 10.3 Hz, 1H), 4.26 (dd, J=17.1, 8.5 Hz, 1H), 3.12-2.97 (m, 1H), 2.97-2.84 (m, 1H), 2.64-2.55 (m, 2H), 2.45-2.34 (m, 2H), 2.16 (s, 3H), 2.10-2.02 (m, 1H), 2.02-1.93 (m, 1H), 1.93-1.81 (m, 1H), 1.74-1.57 (m, 3H), 1.50 (s, 1H), 0.97 (s, 3H).

Example 124: 3-(1-oxo-5-(((1S,2S)-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-144)

Step 1. ((3R,4S)-tetrahydrofuran-3,4-diyl)dimethanol (124-2)

To cis-tetrahydrofuran-3,4-dicarboxylic acid (124-1, 690 mg, 4.31 mmol) in THF (8 mL) at 0° C. and under an atmosphere of nitrogen atmosphere was added BH₃ (17.2 mL, 17.2 mmol) (1M in THF) and the resulting mixture was stirred at rt overnight. The reaction mixture was then quenched with MeOH (2 mL) at 0° C. and concentrated to dryness to afford 124-2 as a colorless oil. The crude product was taken on to the next step without purification. ¹H NMR (400 MHz, Chloroform-d) δ 4.28-4.12 (m, 2H), 4.00-3.87 (m, 2H), 3.82-3.65 (m, 2H), 3.59-3.44 (m, 2H), 1.64-1.48 (m, 2H), 1.48-1.32 (m, 2H).

Step 2. ((3R,4S)-tetrahydrofuran-3,4-diyl)bis(methylene) dimethanesulfonate (124-3)

To the crude ((3R,4S)-tetrahydrofuran-3,4-diyl)dimethanol (124-2, 569 mg, 4.31 mmol) in DIPEA (3.15 mL, 18.1 mmol) and DCM (10 mL) was added methanesulfonyl chloride (0.733 mL, 9.47 mmol) at 0° C. The resulting mixture was stirred at rt for 3 h, then diluted with H₂O and extracted with DCM (×3). The combined organic phases were concentrated to dryness to afford 124-3 (1.21 g, 4.19 mmol, 97% yield) as a brown oil. ¹H NMR (400 MHz, Chloroform-d) δ 4.41-4.34 (m, 2H), 4.32-4.25 (m, 2H), 4.04-3.96 (m, 2H), 3.79-3.71 (m, 2H), 3.08 (s, 6H), 2.92-2.80 (m, 2H).

Step 3. 3-(1-oxo-5-(((1S,2S)-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-144)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 500 mg, 1.46 mmol), DIPEA (1.3 mL, 7.3 mmol) and TBAI (27 mg, 0.073 mmol) in DMF (5 mL) was added ((3R,4S)-tetrahydrofuran-3,4-diyl)bis(methylene) dimethanesulfonate (124-3, 1.05 g, 3.64 mmol) and the resulting mixture was stirred at 80° C. for 55 h. The reaction mixture was then concentrated and the obtained crude material was purified via silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-144 (30.4 mg, 68.0 μmol, 5% yield) as a light beige solid. MS [M+H]⁺=440.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.66 (dd, J=8.6, 4.2 Hz, 1H), 7.00-6.88 (m, 2H), 5.19-4.96 (m, 1H), 4.74-4.57 (m, 1H), 4.38-4.14 (m, 2H), 3.77-3.60 (m, 2H), 3.55-3.41 (m, 2H), 2.89-2.60 (m, 7H), 2.49-2.31 (m, 2H), 2.30-2.18 (m, 1H), 2.16-2.03 (m, 2H), 2.02-1.85 (m, 1H), 1.81-1.65 (m, 3H), 1.62-1.50 (m, 1H).

Example 125: 3-(1-oxo-5-(((1S,2S)-2-((pyridin-2-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-145)

To 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 71.6 mg, 0.184 mmol) and picolinaldehyde (68-1, 22.9 mg, 0.214 mmol) in 2,2,2-trifluoroethanol (1 mL) was added sodium triacetoxyborohydride (55.3 mg, 0.261 mmol) in one portion and the resulting mixture was stirred at rt for 1 h. Additional picolinaldehyde (68-1, 20 mg, 0.19 mmol) and sodium triacetoxyborohydride (39 mg, 0.18 mmol) were added until complete disappearance of I-12 was observed. The reaction mixture was diluted with EtOAc (80 mL) and washed with saturated sodium bicarbonate solution (10 mL) and brine (10 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated to dryness. The crude reaction mixture was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-145 (49.8 mg, 0.103 mmol, 56% yield) as a white solid. MS [M+H]⁺=435.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.53-8.45 (m, 1H), 8.14 (s, 1H), 7.74 (tt, J=7.6, 2.0 Hz, 1H), 7.63-7.57 (m, 1H), 7.47-7.40 (m, 1H), 7.27-7.20 (m, 1H), 7.14 (dd, J=4.1, 2.2 Hz, 1H), 7.06-6.92 (m, 1H), 5.07 (ddd, J=13.3, 5.1, 2.4 Hz, 1H), 4.72-4.64 (m, 1H), 4.45-4.19 (m, 2H), 3.97-3.82 (m, 2H), 3.21-3.15 (m, 2H), 2.91 (ddd, J=18.2, 13.7, 5.4 Hz, 1H), 2.63-2.55 (m, 1H), 2.44-2.34 (m, 1H), 2.21-2.11 (m, 1H), 2.04-1.87 (m, 2H), 1.83-1.60 (m, 3H), 1.59-1.47 (m, 1H).

Example 126: 3-(1-oxo-5-(((1S,2S)-2-(pyrrolidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-146)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 100 mg, 0.291 mmol) in MeCN (3 mL) was added 1,4-dibromobutane (66-1, 0.035 mL, 0.29 mmol) and DIPEA (0.11 mL, 0.64 mmol) and the resulting mixture was stirred at 65° C. overnight. The reaction mixture was cooled to rt and then added to a saturated aqueous solution of sodium hydrogen carbonate (20 mL). The aqueous mixture was extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and the organic solvent concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-146 (47 mg, 12 μmol, 39% yield) as a pale yellow solid. MS [M+H]⁺=398.5. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.15 (s, 1H), 7.03 (d, J=8.4 Hz, 1H), 5.06 (dd, J=12.8, 5.2 Hz, 1H), 4.79-4.68 (m, 1H), 4.44-4.20 (m, 2H), 2.90 (ddd, J=18.0, 13.7, 5.4 Hz, 1H), 2.83-2.70 (s, 1H), 2.65-2.50 (m, 4H, obscured by DMSO-signal), 2.39 (td, J=13.2, 4.4 Hz, 2H), 2.21-2.10 (s, 1H), 2.02-1.88 (d, J=14.4 Hz, 2H), 1.75-1.51 (m, 8H).

Example 127: 3-(5-(((1S,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-147)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 30 mg, 0.087 mmol) in MeCN (2 mL) was added DIPEA (0.076 mL, 0.44 mmol) followed by 3,3-bis(bromomethyl)oxetane (47-1, 64 mg, 0.26 mmol) and the resulting mixture was stirred at 80° C. for 2 days. The reaction mixture was then cooled to rt and additional 3,3-bis(bromomethyl)oxetane (47-1, 64 mg, 0.26 mmol) and DIPEA (0.076 mL, 0.44 mmol) were added. The reaction mixture was stirred at 80° C. for 3 days and then concentrated to dryness, taken up in DCM, and poured into saturated aqueous NaHCO₃. The phases were separated and the aqueous phase was extracted with DCM (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃). The obtained product was taken up in DCM and washed with saturated NaHCO₃ (aq). The phases were separated and the aqueous phase was extracted with DCM (×2). The combined organic phases were concentrated to dryness to afford I-147 (13 mg, 0.027 mmol, 32% yield) a light brown solid. MS [M+H]⁺=426.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.13 (d, J=2.2 Hz, 1H), 7.01 (dt, J=8.6, 1.9 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.60-4.54 (m, 4H), 4.50 (dd, J=6.1, 2.2 Hz, 1H), 4.39 (dd, J=17.2, 2.9 Hz, 1H), 4.26 (d, J=17.1 Hz, 1H), 3.30-3.26 (m, 4H, shoulder on H₂O signal), 2.96-2.84 (m, 1H), 2.78-2.72 (m, 1H), 2.62-2.55 (m, 1H), 2.45-2.35 (m, 1H), 2.12-2.02 (m, 1H), 2.03-1.90 (m, 1H), 1.79-1.52 (m, 4H), 1.40-1.30 (m, 1H).

Example 128: 3-(5-(((1S,2S)-2-(bis((3-methyloxetan-3-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-148)

To a suspension of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 146 mg, 0.130 mmol) in DMF (1 mL) was added 3-methyloxetane-3-carbaldehyde (50-1, 40 mg, 0.39 mmol), followed by sodium triacetoxyborohydride (83 mg, 0.39 mmol). The reaction mixture was stirred at rt for 1 h. Additional 3-methyloxetane-3-carbaldehyde (50-1, 50 mg, 0.49 mmol) and sodium triacetoxyhydroborate (60 mg, 0.28 mmol) were added and stirring was continued at rt for 16 h. Additional 3-methyloxetane-3-carbaldehyde (50-1, 50 mg, 0.490 mmol) and sodium triacetoxyhydroborate (60 mg, 0.28 mmol) were added and stirring was continued at rt for 8 h. The reaction mixture was then was concentrated onto Celite®, azeotroping with heptanes (×2). The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc in heptane to afford I-148 (98.0 mg, 0.186 mmol, 91% yield) as a colorless oil. MS [M+H]⁺=512.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.15 (s, 1H), 7.02 (dd, J=8.3, 2.2 Hz, 1H), 5.06 (dd, J=14.0, 4.4 Hz, 1H), 4.68 (s, 1H), 4.46-4.19 (m, 6H), 4.12 (d, J=5.6 Hz, 4H), 2.88 (t, J=6.2 Hz, 1H), 2.72 (d, J=3.4 Hz, 1H), 2.59 (d, J=13.2 Hz, 4H), 2.45-2.27 (m, 2H), 2.10-1.92 (m, 2H), 1.81 (d, J=8.9 Hz, 1H), 1.73-1.39 (m, 4H), 1.27 (s, 6H).

Example 129: 3-(5-(((1S,2S)-2-(4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-149)

Step 1. 3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (129-2)

To a solution of 3-methylpentane-1,5-diol (129-1, 650 mg, 5.50 mmol) in MeCN (15 mL) was added DMAP (100 mg, 0.819 mmol) and TEA (3.1 mL, 22 mmol), followed by TsCl (3.00 g, 15.7 mmol) and the resulting mixture was stirred at rt for 3 h. The reaction mixture was then filtered and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-50% EtOAc in heptane to afford 129-2 (2.27 g, 5.32 mmol, 97% yield) as a light yellow oil. MS [M+H₂O]⁺=444.5. ¹H NMR (400 MHz, Chloroform-d) δ 7.80-7.76 (m, 4H), 7.35 (dd, J=8.7, 0.7 Hz, 4H), 4.05-3.97 (m, 4H), 2.46 (s, 6H), 1.70-1.58 (m, 3H), 1.49-1.36 (m, 2H), 0.78 (d, J=6.5 Hz, 3H).

Step 2. 3-(5-(((1S,2S)-2-(4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-149)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 75.0 mg, 0.218 mmol) in MeCN (2 mL) was added DIPEA (0.23 mL, 1.3 mmol), followed by 3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (129-2, 466 mg, 1.09 mmol) and the resulting solution was stirred at 120° C. under μW irradiation for 1 h. The reaction mixture was then concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-10% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃). The obtained material was taken up in DCM and washed with saturated NaHCO₃ (aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic phases were concentrated to dryness to afford I-149 (36.0 mg, 0.080 mmol, 37% yield) as an off-white solid. MS [M+H]⁺=426.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.2 Hz, 1H), 7.03 (d, J=8.3 Hz, 1H), 5.06 (dd, J=13.2, 5.0 Hz, 1H), 4.73-4.66 (m, 1H), 4.45-4.18 (m, 2H), 2.97-2.76 (m, 3H), 2.63-2.55 (m, 1H), 2.44-2.35 (m, 1H), 2.10-1.83 (m, 5H), 1.75-1.42 (m, 5H), 1.37-1.19 (m, 2H), 1.17-1.01 (m, 2H), 0.99-0.92 (m, 1H), 0.86 (d, J=6.5 Hz, 3H).

Example 130: 3-(1-oxo-5-(((1S,2S)-2-((pyridin-3-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-150)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 100 mg, 0.291 mmol) in MeCN (3 mL) was added 3-(bromomethyl)pyridine HBr (130-1, 73.7 mg, 0.291 mmol) and DIPEA (0.11 mL, 0.64 mmol) and the resulting mixture was stirred at 65° C. for 3 hours. The reaction mixture was cooled to rt and then added to a saturated aqueous solution of sodium hydrogen carbonate (20 mL). The aqueous mixture was extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and the organic solvent concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃). The obtained material was treated with 1:1 MeCN/diethyl ether. The resulting suspension was sonicated and filtered and the obtained solid was washed with diethyl ether. The filtrate was collected and concentrated to dryness. The material was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-150 (8.0 mg, 0.016 mmol, 5% yield). MS [M+H]⁺=435.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 8.55 (s, 1H), 8.43 (d, J=4.8 Hz, 1H), 8.12 (s, 1H), 7.76 (d, J=7.9 Hz, 1H), 7.59 (d, J=8.4 Hz, 1H), 7.33 (dd, J=7.8, 4.8 Hz, 1H), 7.14 (s, 1H), 7.04-6.96 (m, 1H), 5.11-5.02 (m, 1H), 4.69-4.64 (m, 1H), 4.42-4.21 (m, 2H), 3.87-3.75 (m, 2H), 3.18-3.12 (m, 2H), 2.91 (ddd, J=17.7, 13.6, 5.4 Hz, 1H), 2.60 (d, J=17.7 Hz, 1H), 2.45-2.32 (m, 1H), 2.20-2.09 (m, 1H), 2.02-1.88 (m, 2H), 1.80-1.60 (multiplets, 3H), 1.58-1.48 (m, 1H).

Example 131: 3-(1-oxo-5-(((1S,2S)-2-((pyridin-4-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-151)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 100 mg, 0.291 mmol) in TFE (3 mL) was added isonicotinaldehyde (57-1, 31 mg, 0.29 mmol) and the resulting mixture was stirred at rt for 15 minutes. Sodium triacetoxyborohydride (93 mg, 0.44 mmol) was added and stirring was continued at rt for 3 hours. The reaction mixture was then added to a saturated aqueous solution of sodium hydrogen carbonate (20 mL). The aqueous mixture was extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and the organic solvent evaporated concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) and then further purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford crude material. The obtained material was then again purified via silica gel chromatography eluting with 0-20% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-151 (10.0 mg, 0.022 mmol, 8% yield) as a white solid. MS [M+H]⁺=435.5. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm); 10.96 (s, 1H), 8.56-8.43 (m, 2H), 7.60 (d, J=8.4 Hz, 1H), 7.45-7.33 (m, 2H), 7.15 (d, J=8.0 Hz, 1H), 7.05-6.98 (m, 1H), 5.12-5.02 (m, 1H), 4.72-4.57 (br m, 1H), 4.43-4.20 (m, 2H), 3.78 (br s, 2H), 3.14-3.04 (m, 1H), 2.99-2.82 (m, 1H), 2.60 (d, J=17.6 Hz, 2H), 2.39 (br d, J=12.3 Hz, 1H), 2.23-2.10 (br m, 1H), 2.04-1.86 (m, 2H), 1.84-1.45 (br m, 4H).

Example 132: 3-(5-(((1S,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-152)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 50.0 mg, 0.146 mmol) in MeCN (1.5 mL) was added DIPEA (0.13 mL, 0.73 mmol), followed by 3-methoxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (44-3, 165 mg, 0.361 mmol) and the resulting solution was stirred at 120° C. under μW irradiation for 1 h. The reaction mixture was then concentrated to dryness and the crude residue was taken up in DCM and washed with saturated NaHCO₃ (aq) and the phases were separated. The aqueous layer was extracted with DCM and the combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-10% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-152 (22.0 mg, 0.046 mmol, 32% yield) as an off-white solid. MS [M+H]⁺=456.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.2 Hz, 1H), 7.03 (d, J=8.4 Hz, 1H), 5.07 (dd, J=13.4, 5.0 Hz, 1H), 4.69 (s, 1H), 4.47-4.17 (m, 2H), 3.06 (s, 3H), 2.97-2.82 (m, 2H), 2.59 (d, J=16.9 Hz, 1H), 2.45-2.28 (m, 5H), 2.11-1.85 (m, 3H), 1.72-1.55 (m, 5H), 1.43 (s, 3H), 1.06 (s, 3H).

Example 133: 3-(5-(((1S,2S)-2-(4,4-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-153) Step 1. 3,3-dimethylpentane-1,5-diol (133-2)

To a solution of 3,3-dimethylpentanedioic acid (133-1, 600 mg, 3.75 mmol) in THF (15 mL) cooled to −20° C. was slowly added BH₃ (1M in THF) (9.4 mL, 9.4 mmol) and the resulting mixture was stirred at −20° C., then warmed to rt and stirred at rt overnight. The reaction mixture was cooled to 0° C. and quenched with water (8 mL). K₂CO₃ was added and the aqueous mixture was extracted with Et₂O (×3). The combined organic phases were concentrated to dryness to afford crude 133-2 (516 mg) as a light yellow oil. The product was taken onto the next step without purification. ¹H NMR (400 MHz, Chloroform-d) δ 3.75 (t, J=7.0 Hz, 4H), 1.58 (t, J=7.0 Hz, 4H), 0.95 (s, 6H).

Step 2. 3,3-dimethylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (133-3)

To a solution of 3,3-dimethylpentane-1,5-diol (133-2, 516 mg, 3.90 mmol) in MeCN (20 mL) was added DMAP (238 mg, 1.95 mmol) and TEA (2.2 mL, 16 mmol), followed by TsCl (1.86 g, 9.76 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was then filtered and concentrated to dryness. The crude material was purified via silica gel chromatography eluting 0-50% EtOAc in heptane to afford 133-3 (860 mg, 1.95 mmol, 50% yield) as a pale yellow solid. MS [M+H]⁺=441.5. ¹H NMR (400 MHz, Chloroform-d) δ 7.80-7.75 (m, 4H), 7.38-7.33 (m, 4H), 4.03 (t, J=7.1 Hz, 4H), 2.46 (s, 6H), 1.56 (t, J=7.1 Hz, 4H), 0.85 (s, 6H).

Step 3. 3-(5-(((1S,2S)-2-(4,4-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-153)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 50.0 mg, 0.146 mmol) in MeCN (2 mL) was added DIPEA (0.13 mL, 0.73 mmol), followed by 3,3-dimethylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (133-3, 96.0 mg, 0.218 mmol) and the resulting solution was stirred at 120° C. under μW irradiation for 3 h. The reaction mixture was then concentrated to dryness. The crude material was taken up in DCM and washed with saturated aqueous NaHCO₃. The phases were separated and the aqueous phase was extracted with DCM and the combined organic phases were dried over Na₂SO₄, filtered, and concentrated. The crude product was purified via silica gel chromatography eluting with 0-10% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-153 (26 mg, 0.056 mmol, 39% yield) as an off-white solid. MS [M+H]⁺=440.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.1 Hz, 1H), 7.03 (d, J=8.2 Hz, 1H), 5.06 (dd, J=13.2, 5.1 Hz, 1H), 4.70 (s, 1H), 4.39 (dd, J=17.1, 9.9 Hz, 1H), 4.26 (dd, J=17.1, 8.1 Hz, 1H), 2.96-2.81 (m, 2H), 2.65-2.55 (m, 1H), 2.47-2.30 (m, 5H), 2.12-1.86 (m, 3H), 1.72-1.55 (m, 3H), 1.56-1.41 (m, 1H), 1.37-1.23 (m, 4H), 0.88 (s, 6H).

Example 134: 3-(5-(((1S,2S)-2-(4-methoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-154)

Step 1. Diethyl 3-methoxypentanedioate (134-2)

To a mixture of diethyl 3-hydroxygluturate (134-1, 0.455 mL, 2.45 mmol) and silver oxide (1.42 g, 6.12 mmol) in DMF (5 mL), was added methyl iodide (0.77 mL, 12 mmol) dropwise during which the reaction temperature was maintained at 25-30° C. with a water bath. The resulting mixture was then stirred at rt over 3 days. The reaction mixture was filtered and the formed precipitate was washed with diethyl ether. The filtrate was washed with 5% NaHSO₃ (aq) and brine and the organic phase was dried over Na₂SO₄, filtered, and concentrated to dryness to afford 134-2 (440 mg, 2.02 mmol, 82% yield) as a colorless oil. ¹H NMR (400 MHz, Chloroform-d) δ 4.25-4.13 (m, 4H), 4.13-4.05 (m, 1H), 3.41 (s, 3H), 2.71-2.51 (m, 4H), 1.36-1.21 (m, 6H).

Step 2. 3-methoxypentane-1,5-diol (134-3)

To a solution of diethyl 3-methoxypentanedioate (134-2, 250 mg, 1.15 mmol) in THF (20 mL) cooled to 0° C. was slowly added 1M lithium aluminum hydride in THF (3.44 mL, 3.44 mmol) and the resulting mixture was stirred at 0° C. and then warmed to rt and stirred at rt for 3 h. The reaction mixture was cooled to 0° C., quenched with saturated Na₂SO₄ (aq), and filtered. The obtained precipitate was washed with diethyl ether. The filtrate was washed with 5% NaHSO₃ (aq) and brine, dried over Na₂SO₄, filtered, and concentrated to dryness to afford 134-3 which was taken on to the next step without purification.

Step 3. 3-methoxypentane-1,5-diyl bis(4-methylbenzenesulfonate) (134-4)

To a solution of 3-methoxypentane-1,5-diol (134-3, 92.0 mg, 0.686 mmol) in MeCN (5 mL) was added DMAP (42 mg, 0.34 mmol) and TEA (0.38 mL, 2.7 mmol), followed by TsCl (327 mg, 1.71 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was then filtered and concentrated to dryness. The obtained residue was treated with diethyl ether and filtered. The filtrate was concentrated to dryness and the crude material was purified via silica gel chromatography eluting with 0-60% EtOAc in heptane to afford 134-4 (40.0 mg, 0.090 mmol, 13% yield) a light yellow oil. MS [M+H₂O]⁺=460.5. ¹H NMR (400 MHz, Chloroform-d) δ 7.81-7.76 (m, 4H), 7.38-7.32 (m, 4H), 4.13-4.01 (m, 4H), 3.42-3.32 (m, 1H), 3.15 (s, 3H), 2.45 (s, 6H), 1.84-1.68 (m, 4H).

Step 4. 3-(5-(((1S,2S)-2-(4-methoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-154)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 31 mg, 0.090 mmol) in MeCN (1 mL) was added DIPEA (0.047 mL, 0.27 mmol), followed by 3-methoxypentane-1,5-diyl bis(4-methylbenzenesulfonate) (134-4, 40 mg, 0.090 mmol) and the resulting solution was stirred at 120° C. under μW irradiation for 4 h. The reaction mixture was concentrated to dryness and the crude residue was taken up in DCM and washed with saturated NaHCO₃(aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-10% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-154 (11 mg, 0.022 mmol, 25% yield) as an off-white solid. MS [M+H]⁺=442.6. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (d, J=1.7 Hz, 1H), 7.03 (d, J=8.6 Hz, 1H), 5.07 (dd, J=13.2, 5.0 Hz, 1H), 4.73-4.66 (m, 1H), 4.45-4.18 (m, 2H), 3.20 (s, 3H), 3.17-3.09 (m, 1H), 2.97-2.81 (m, 2H), 2.79-2.66 (m, 1H), 2.59 (d, J=17.0 Hz, 1H), 2.44-2.34 (m, 1H), 2.22-1.86 (m, 5H), 1.86-1.74 (m, 2H), 1.72-1.54 (m, 4H), 1.53-1.31 (m, 3H).

Example 135: 3-(5-(((1S,2S)-2-(ethyl(oxetan-3-ylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-155)

To a solution of 3-(5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-29, 0.18 g, 0.49 mmol) and oxetane-3-carbaldehyde (135-1, 60 mg, 0.73 mmol) in DCE (10 mL) at 0° C. was added NaBH(OAc)₃ (0.15 g, 0.73 mmol) and the resulting mixture was stirred at rt for 16 h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (2×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified via silica gel chromatography eluting with 5% MeOH in DCM to afford compound I-155 (70 mg, 0.16 mmol, 33% yield) as an off-white solid. MS [M+H]⁺=442.2. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.61 (d, J=8.8 Hz, 1H), 7.22 (s, 1H), 7.06 (d, J=8.8 Hz, 1H), 5.10-5.05 (m, 1H), 4.66-4.62 (m, 1H), 4.57-4.53 (m, 2H), 4.37-4.25 (m, 2H), 4.21-4.18 (m, 2H), 3.22-3.12 (m, 2H), 2.92-2.89 (m, 1H), 2.75-2.72 (m, 2H), 2.60-2.55 (m, 1H), 2.46-2.32 (m, 3H), 2.05-1.95 (m, 2H), 1.90-1.88 (m, 1H), 1.66-1.62 (m, 3H), 1.52-1.49 (m, 1H), 0.96-0.93 (m, 3H).

Example 136: 3-(5-(((1S,2S)-2-(isoindolin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-156)

To a stirred solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 200 mg, 0.580 mmol) and DIPEA (0.300 mL, 1.74 mmol) in DMF (10 mL) was added 1,2-bis(bromomethyl)benzene (136-1, 230 mg, 0.860 mmol), followed by TBAI (20 mg, 0.060 mmol) and the resulting mixture was stirred at 60° C. for 16 h. The reaction mixture was then cooled to rt, diluted with EtOAc. The organic phase was washed with saturated aqueous NaHCO₃ and brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified via silica gel chromatography eluting with 10% MeOH in DCM and then further purified by reverse phase HPLC (Column: ZOBRAX ECLIPS XDB C18 (150×21.5 μm), Mobile phase A: 0.01% TFA (aq), Mobile phase B: MeCN, Time (min)/% B: 0/25, 2/35, 9/65, Flow rate: 18 ml/min) to afford I-156 (15 mg, 0.03 mmol, 6% yield) as a white solid. MS [M+H]⁺=446.2. ¹H NMR (400 MHz, DMSO-d₆): δ 10.98 (s, 1H), 7.63 (d, J=8.8 Hz, 1H), 7.23-7.15 (m, 5H), 7.08 (d, J=8.8 Hz, 1H), 5.10-5.06 (m, 1H), 4.86-4.84 (m, 1H), 4.42 (d, J=17.2, 1H), 4.28 (d, J=17.2, 1H), 3.96-3.87 (m, 3H) 3.20-3.16 (m, 1H), 2.95-2.86 (m, 1H), 2.67-2.61 (m, 1H), 2.40-2.32 (m, 2H), 2.21-2.10 (m, 1H), 2.08-1.96 (m, 2H), 1.75-1.65 (m, 4H).

Example 137: 3-(5-(((1S,2S)-2-(3-methoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-157)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 50.0 mg, 0.146 mmol) in MeCN (1.5 mL) was added DIPEA (0.15 mL, 0.87 mmol), followed by 2-methoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) (52-3, 177 mg, 0.427 mmol) and the resulting solution was stirred at 120° C. under μW irradiation for 4 h. The reaction mixture was concentrated to dryness and the crude residue was taken up in DCM and washed with saturated NaHCO₃ (aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-157 (13.0 mg, 0.028 mmol, 19% yield) as clear glassy solid. MS [M+H]⁺=414.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.14 (d, J=2.1 Hz, 1H), 7.02 (dd, J=8.3, 2.2 Hz, 1H), 5.75 (s, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.56-4.50 (m, 1H), 4.39 (dd, J=17.2, 3.8 Hz, 1H), 4.26 (dd, J=17.1, 3.6 Hz, 1H), 3.97-3.87 (m, 1H), 3.54-3.44 (m, 2H), 3.13 (s, 3H), 2.89-2.78 (m, 3H), 2.63-2.55 (m, 1H), 2.43-2.35 (m, 1H), 2.15-2.04 (m, 1H), 2.03-1.94 (m, 1H), 1.82-1.72 (m, 1H), 1.72-1.55 (m, 3H), 1.44-1.35 (m, 1H).

Example 138: 3-(5-(((1S,2S)-2-(4-ethoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-158)

Step 1. 3-ethoxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (138-1)

To a solution of 3-hydroxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (44-2, 250 mg, 0.565 mmol) and Proton-sponge® (242 mg, 1.13 mmol) in DCM was added triethyloxonium tetrafluoroborate (1M in DCM) (1.1 mL, 1.1 mmol) and the resulting mixture was stirred at rt overnight. Additional triethyloxonium tetrafluoroborate (1M in DCM) (1.1 mL, 1.1 mmol) was added and stirring was continued at rt overnight. The reaction mixture was then filtered and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-60% EtOAc in heptane to afford 138-1 (61.0 mg, 0.130 mmol, 23% yield) as a colorless oil. MS [M+H₂O]⁺=488.4. ¹H NMR (400 MHz, Chloroform-d) δ 7.80-7.75 (m, 4H), 7.37-7.33 (m, 4H), 4.11-4.01 (m, 4H), 3.18 (q, J=7.0 Hz, 2H), 2.45 (s, 6H), 1.88-1.72 (m, 4H), 1.08 (s, 3H), 1.00 (t, J=6.9 Hz, 3H).

Step 2. 3-(5-(((1S,2S)-2-(4-ethoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-158)

To a solution of 3-ethoxy-3-methylpentane-1,5-diyl bis(4-methylbenzenesulfonate) (138-1, 61 mg, 0.13 mmol) and 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 40 mg, 0.12 mmol) in MeCN (1 mL) was added DIPEA (0.10 mL, 0.58 mmol) and the resulting solution was stirred at 120° C. under μW irradiation for 12 h. The reaction mixture was then concentrated to dryness and the crude residue was taken up in DCM and washed with saturated NaHCO₃ (aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-158 (10 mg, 0.019 mmol, 16% yield) as clear glassy solid. MS [M+H]⁺=470.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.20-7.14 (m, 1H), 7.03 (d, J=8.3 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.73-4.65 (m, 1H), 4.43-4.19 (m, 2H), 3.30-3.24 (m, 3H, shoulder on H₂O signal), 2.98-2.81 (m, 2H), 2.64-2.54 (m, 1H), 2.47-2.34 (m, 4H), 2.11-1.88 (m, 3H), 1.73-1.55 (m, 5H), 1.54-1.35 (m, 3H), 1.10-1.00 (m, 6H).

Example 139: 3-(5-(((1S,2S)-2-((((1r,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-159)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 0.20 g, 0.53 mmol) and (1r,4r)-4-methoxycyclohexane-1-carbaldehyde (48-3, 0.15 g, 1.1 mmol) in TFE (10 mL) was added NaBH(OAc)₃ (0.17 g, 0.78 mmol) at 0° C. and the resulting mixture was stirred at rt for 16 h. The reaction mixture was quenched with water and extracted with DCM (3×50 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified via silica gel chromatography eluting with 5% MeOH in DCM to afford I-159 (22 mg, 0.050 mmol, 10% yield) as a pale green solid. [M+H]⁺=470.3. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.60 (d, J=8.8 Hz, 1H), 7.17 (s, 1H), 7.04 (d, J=8.8 Hz, 1H), 5.12-5.05 (m, 1H), 4.58-4.52 (m, 1H), 4.38-4.25 (m, 2H), 3.20 (s, 3H), 3.19-2.87 (m, 3H), 2.62-2.51 (m, 2H), 2.38-2.32 (m, 2H), 2.13-2.09 (m, 1H), 1.98-1.95 (m, 4H), 1.79-1.62 (m, 5H), 1.42-1.28 (m, 3H), 1.05-0.72 (m, 4H).

Example 140: (1S,4r)-4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)amino)methyl)cyclohexane-1-carbonitrile (I-160)

Step 1. Methyl 4-carbamoylcyclohexane-1-carboxylate (140-2)

To a cooled solution of (1r,4r)-4-(methoxycarbonyl)cyclohexane-1-carboxylic acid (140-1, 5.00 g, 15.2 mmol) in tetrahydrofuran (100 mL) at −10° C. was sequentially added TEA (7.2 mL, 38 mmol) and ethyl chloroformate (2.25 mL, 22.9 mmol) under an atmosphere of nitrogen and the resulting mixture was stirred at room temperature for 3 h. The reaction mixture was then cooled to −10° C., ammonium hydroxide (25.0 mL, 91.2 mmol) was added and the mixture warmed to room temperature and stirred overnight. The reaction mixture was diluted with water and extracted with EtOAc (3×50 mL). The combined organic phases were washed with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated to afford 140-2 (4.80 g, 25.9 mmol, 96% yield) as a white solid. ¹H NMR (400 MHz, DMSO-d₆): δ 7.18 (br s, 1H), 6.68 (br s, 1H), 3.58 (s, 3H), 2.28-2.20 (m, 1H), 2.18-1.98 (m, 1H), 1.92-1.86 (m, 2H), 1.78-1.74 (m, 2H), 1.35-1.30 (m, 4H).

Step 2. Methyl 4-cyanocyclohexane-1-carboxylate (140-3)

To a solution of the methyl 4-carbamoylcyclohexane-1-carboxylate (140-2, 2.80 g, 11.4 mmol) in pyridine (28.0 mL) was added imidazole (0.790 g, 11.4 mmol) and phosphorous oxychloride (2.80 mL) in one portion at 0° C. under an atmosphere of nitrogen and the resulting mixture was stirred at room temperature for 4 h. The reaction mixture was then quenched with water and extracted with EtOAc (3×50 mL). The combined organic phases were washed with 2 M hydrochloric acid (aq), dried over Na₂SO₄, filtered, and concentrated. The obtained crude material was purified via silica gel chromatography eluting with 30% EtOAc in hexanes to afford 140-3 (1.60 g, 9.58 mmol, 64% yield) as a white solid. ¹H NMR (300 MHz, CDCl₃): δ 3.67 (s, 3H), 2.48-2.33 (m, 2H), 2.16-2.02 (m, 4H), 1.69-1.48 (m, 4H).

Step 3. 4-(hydroxymethyl)cyclohexane-1-carbonitrile (140-4)

To a solution of methyl 4-cyanocyclohexane-1-carboxylate (140-3, 1.00 g, 5.90 mmol) in MeOH (10 mL) at 0° C. was added NaBH₄ (0.540 g, 14.8 mmol) in small portions at 0° C. and the resulting mixture was allowed to warm to rt and stirred at rt for 6 h. The reaction mixture was then quenched with ice-cold water and extracted with EtOAc (3×50 mL). The combined organic phases were washed with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated. The obtained crude material was purified via silica gel chromatography eluting with 30% EtOAc in hexanes to afford 140-4 (0.520 g, 3.74 mmol, 63% yield) as an off-white solid. ¹H NMR (400 MHz, CDCl₃): δ 3.49-3.47 (m, 2H), 2.41-2.34 (m, 1H), 2.18-2.11 (m, 2H), 1.91-1.86 (m, 2H), 1.63-1.49 (m, 3H), 1.39-1.36 (m, 1H), 1.05-0.95 (m, 2H).

Step 4. 4-formylcyclohexane-1-carbonitrile (140-5)

To a solution of 4-(hydroxymethyl)cyclohexane-1-carbonitrile (140-4, 0.400 g, 2.87 mmol) in DCM (20 mL) at 0° C. was added PCC (1.23 g, 5.74 mmol) and the resulting mixture was stirred at rt for 6 h. The reaction mixture was then filtered through Celite® and washed with DCM. The filtrate was concentrated to dryness and the crude material was purified via silica gel chromatography eluting with 40% EtOAc in hexanes to afford 140-5 (0.150 g, 1.09 mmol, 38%) as pale yellow oil. ¹H NMR (400 MHz, CDCl₃): δ 9.64 (s, 1H), 2.51-2.33 (m, 2H), 2.11-2.06 (m, 4H), 1.69-1.64 (m, 2H), 1.50-1.45 (m, 2H).

Step 5. (1S,4r)-4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)amino)methyl)cyclohexane-1-carbonitrile (I-160)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (I-12, 0.250 g, 0.658 mmol) and (1r,4r)-4-formylcyclohexane-1-carbonitrile (140-5, 0.135 g, 0.987 mmol) in DCE (10 mL) at 0° C. was added NaBH(OAc)₃ (0.418 g, 1.97 mmol) and the resulting mixture was stirred at rt for 16 h. The reaction mixture was then quenched with saturated aqueous sodium bicarbonate solution and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified by reverse phase HPLC (Column: KINETEX EVO C18, (21.2 mm×150 mm), Mobile phase: A:0.01% ammonia, B: acetonitrile, Flow rate: 18 ml/min, Time (min)/% B 0/20, 2/30, 8/50) to afford I-160 (0.020 g, 0.172 mmol, 26% yield) as a white solid. MS [M+H]⁺=465.15. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.60 (d, J=8.8 Hz, 1H), 7.16 (s, 1H), 7.03 (d, J=8.8 Hz, 1H), 5.10-5.05 (m, 1H), 4.54-4.52 (m, 1H), 4.36-4.25 (m, 2H), 3.06-3.02 (m, 1H), 2.90-2.88 (m, 1H), 2.66-2.55 (m, 2H), 2.38-2.33 (m, 4H), 2.07-1.85 (m, 4H), 1.78-1.61 (m, 6H), 1.44-1.37 (m, 4H), 0.91-0.88 (m, 2H).

Example 141: 3-(5-(((1S,2S)-2-(((4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione TFA Salt (I-161)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (I-12, 0.200 g, 0.526 mmol) and 4-methoxycyclohexane-1-carbaldehyde (141-1, 0.112 g, 0.782 mmol) in TFE (10 mL) at 0° C. was added NaBH(OAc)₃ (0.167 g, 0.782 mmol) and the resulting mixture was stirred at rt for 2 h. The reaction mixture was then concentrated to dryness and the obtained residue was diluted with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by reverse phase HPLC (Column: KINETEX EVO C18 (150 mm×21.0 mm), 5.0g, Mobile phase: 0.05% TFA in water (A): MeCN (B); Flow: 20 mL/min; Time (min)/% B: 0/10, 2/20, 10/50) to afford the TFA salt of I-161 (0.013 mg, 0.027 mmol, 5% yield) as pale green solid. MS [M+H]⁺=470.3. ¹H NMR (400 MHz, DMSO-d₆): δ 10.99 (s, 1H), 8.79 (br s, 2H), 7.68 (d, J=8.4 Hz, 1H), 7.17 (s, 1H), 7.07 (d, J=8.8 Hz, 1H), 5.08 (dd, J=13.2, 4.8 Hz, 1H), 4.96-4.94 (m, 1H), 4.40-4.24 (m, 2H), 3.71-6.62 (m, 4H), 3.21 (s, 3H), 3.08-3.04 (m, 1H), 2.95-2.86 (m, 3H), 2.55-2.35 (m, 2H), 2.22-2.15 (m, 2H), 2.02-1.95 (m, 3H), 1.80-1.62 (m, 5H), 1.07-0.96 (m, 4H).

z

A 2-5 mL μW vial was charged with 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 100 mg, 0.291 mmol), DIPEA (0.25 mL, 1.5 mmol), oxetane-3,3-diylbis(ethane-2,1-diyl) bis(4-methylbenzenesulfonate) (87-6, 139 mg, 0.306 mmol) and MeCN (1 mL). The resulting mixture was stirred for 4 hours at 120° C. in the μW and then concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collected fractions contained ˜1 drop of formic acid) and then further purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-162 (2.0 mg, 4.7 μmol, 1% yield) as a white solid. MS [M+H]⁺=454.5. ¹H NMR (400 MHz, Deuterium Oxide) δ 8.45 (s, 1H), 7.77 (d, J=8.5 Hz, 1H), 7.22-7.09 (m, 2H), 5.14 (ddd, J=13.3, 5.3, 2.1 Hz, 1H), 5.05-4.95 (m, 1H), 4.64-4.44 (m, 6H), 3.87-3.71 (m, 1H), 3.43-3.06 (m, 3H), 3.00-2.84 (m, 2H), 2.53 (qd, J=12.9, 5.4 Hz, 1H), 2.40-2.22 (m, 4H), 2.21-2.03 (m, 3H), 1.92-1.72 (m, 5H).

Example 143: 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile HC(O)OH Salt (I-163) and (1R,3S)-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile and (1S,3R)-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile (I-164 and I-165)

To a solution of 3-(5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-29, 91 mg, 0.25 mmol) in TFE (1 mL) was added 3-formyl-1-methylcyclobutane-1-carbonitrile (77-4, 91 mg, 0.74 mmol), followed by sodium triacetoxyborohydride (156 mg, 0.735 mmol) and the resulting mixture was stirred at rt overnight. Additional 3-formyl-1-methylcyclobutane-1-carbonitrile (77-4, 120 mg) and sodium triacetoxyborohydride (170 mg, 0.80 mmol) was added until complete disappearance of I-29 was observed. The reaction mixture was then concentrated to dryness and the crude material was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford the formate salt of I-163 (38 mg, 0.078 mmol, 32% yield) as a white solid, as a mixture of diastereoisomers. MS [M+H]⁺=479.5. (3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile HC(O)OH salt (I-163, 34 mg) was purified using chiral SFC (Column: Chiralpak IH 21×250 mm; Flow rate: 80 g per minute; Cosolvent: 30% i-PrOH with 10 mM NH₃) followed by purification via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford two diastereomeric products I-164 and I-165.

Peak 1: Isolated diastereomer I-164 (7 mg, 0.014 mmol, 21% yield) as a white solid. Chiral SFC: Rt=2.25 mins. MS [M+H]⁺=479.2. ¹H NMR (400 MHz, Methylene Chloride-d) δ 8.02 (s, 1H), 7.72 (d, J=8.5 Hz, 1H), 7.11-6.93 (m, 2H), 5.25-5.05 (m, 1H), 4.49-4.27 (m, 2H), 3.66-3.32 (m, 1H), 3.22-2.73 (m, 5H), 2.72-2.40 (m, 2H), 2.41-1.93 (m, 7H), 1.96-1.63 (m, 5H), 1.62-1.33 (m, 3H), 1.28-0.94 (m, 3H). Peak 2: Isolated diastereomer I-165 (3 mg, 6 μmol, 9% yield) as a white solid. Chiral SFC Rt=2.65 mins. MS [M+H]⁺=479.5. ¹H NMR (400 MHz, Methylene Chloride-d) δ 8.06 (s, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.16-6.93 (m, 2H), 5.12 (dd, J=13.2, 5.2 Hz, 1H), 4.50-4.26 (m, 2H), 3.65-3.35 (m, 1H), 3.32-2.68 (m, 6H), 2.73-2.45 (m, 2H), 2.41-2.02 (m, 6H), 1.96-1.64 (m, 5H), 1.54-1.36 (m, 3H), 1.28-0.97 (m, 3H). Relative stereochemistry of the substituents on the cyclobutane ring was not determined and assigned arbitrarily.

Example 144: 3-(1-oxo-5-(((1S,2S)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-206)

Step 1. 2-(2,2,2-trifluoroethoxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (184-1)

To a solution of 106-6 (0.7 g, 4.0 mmol) in DCM (30 mL) was added Et₃N (2.2 mL, 16.1 mmol), DMAP (0.017 g, 0.1 mmol) and TsCl (1.68 g, 8.8 mmol) at 0° C. and resulting mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with cold water and extracted with DCM (3×30 mL). The combined organic phases were washed water, brine, dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 10% EtOAc in hexanes to afford 184-1 (0.83 g, 1.7 mmol, 43%) as pale yellow liquid. MS [M+H+18]⁺=500.1. ¹H NMR (300 MHz, Chloroform-d) δ 7.68 (d, J=8.1 Hz, 4H), 7.37 (d, J=8.1 Hz, 4H), 4.05 (d, J=5.1 Hz, 4H), 3.87-3.84 (m, 3H), 2.46 (s, 6H).

Step 2. 3-(1-oxo-5-(((1S,2S)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-206)

To a solution of I-12 (300 mg, 0.56 mmol) in acetonitrile (10 mL) was added 184-1 (631 mg, 1.31 mmol) and DIPEA (0.77 mL, 4.4 mmol) and resulting mixture was stirred at 120° C. for 9 h under microwave irradiation. Upon complete consumption of starting material, the reaction mixture was diluted with sat. sodium bicarbonate solution and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified by silica gel chromatography eluting with 10% MeOH in DCM to afford compound I-206 (33 mg, 0.06 mmol, 8% yield) as an off-white solid. MS [M+H]⁺=482.4. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.14 (s, 1H), 7.03 (d, J=8.4 Hz, 1H), 5.08-5.04 (m, 1H), 4.58-4.54 (m, 1H), 4.36-4.35 (m, 1H), 4.27-4.20 (m, 2H), 4.05-3.98 (m, 2H), 3.52-3.49 (m, 2H), 2.95-2.87 (m, 4H), 2.66-2.60 (m, 1H), 2.49-2.48 (m, 1H), 2.55-2.45 (m, 2H), 1.80-1.63 (m, 4H), 1.37-1.31 (m, 1H).

Example 145: 3-(5-(((1S,2S)-2-(4-fluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-166)

To a solution of 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-96, 20 mg, 0.047 mmol) in DCM (2 mL) and cooled to 0° C. was added a solution of DAST (17 mg, 0.11 mmol) in DCM (0.2 mL) and the resulting mixture was stirred at 0° C. for 1 h, and then warmed to rt. Upon complete consumption of starting material, the reaction mixture was quenched with saturated NaHCO₃(aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) and then further purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-166 (2.0 mg, 4.7 μmol, 10% yield) as a white solid. MS [M+H]⁺=430.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.19 (d, J=2.2 Hz, 1H), 7.06-7.01 (m, 1H), 5.06 (dd, J=13.1, 5.0 Hz, 1H), 4.76-4.67 (m, 2H), 4.39 (dd, J=17.1, 9.8 Hz, 1H), 4.26 (dd, J=17.1, 8.5 Hz, 1H), 2.96-2.84 (m, 2H), 2.65-2.55 (m, 3H), 2.43-2.34 (m, 3H), 2.12-2.01 (m, 1H), 2.01-1.74 (m, 4H), 1.76-1.55 (m, 5H), 1.55-1.41 (m, 1H).

Example 146: 3-(5-(((1S,2S)-2-(1,5-oxazocan-5-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-167)

Step 1. Dipropyl 3,3′-oxydipropanoate (146-2)

A suspension of p-toluenesulfonic acid (61.3 g, 322 mmol) in n-propanol (100 mL) was added 2-cyanoethyl ether (146-1, 10.0 g, 9.62 mmol) and the resulting mixture was stirred at reflux overnight. The reaction mixture was then cooled to rt, concentrated to roughly 50 mL, and treated with water and heptanes. The phases were separated and the organic phase was washed with aqueous NaHCO₃ and brine. The organic phase was dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc in heptanes to afford 146-2 (11.6 g, 47.1 mmol, 59% yield). ¹H NMR (400 MHz, Chloroform-d) δ 4.07 (t, J=6.7 Hz, 4H), 3.75 (t, J=6.5 Hz, 4H), 2.59 (t, J=6.5 Hz, 4H), 1.66 (quint, J=7.0 Hz, 4H), 0.96 (t, J=7.4 Hz, 6H).

Step 2. 3,3′-oxybis(propan-1-ol) (146-3)

To a solution of dipropyl 3,3′-oxydipropanoate (146-2, 500 mg, 2.03 mmol) in THF (10 mL) at 0° C. was added lithium aluminum hydride (1M in THF) (3.05 mL, 3.05 mmol) and the resulting mixture was stirred at 0° C. and then at rt. Upon complete consumption of starting material, the reaction mixture was cooled to 0° C., slowly quenched with 10% H₂SO₄ (aq) (0.5 mL), and stirred at rt overnight. The organic phase was isolated and then dried over Na₂SO₄, filtered, and concentrated to dryness to afford 146-3 (269 mg, 2.00 mmol, 99% yield) as a colorless oil. ¹H NMR (400 MHz, Chloroform-d) δ 3.74-3.59 (m, 4H), 3.58-3.49 (m, 4H), 1.84-1.66 (m, 4H).

Step 3. Oxybis(propane-3,1-diyl) bis(4-methylbenzenesulfonate) (146-4)

To a solution of 3,3′-oxybis(propan-1-ol) (146-3, 269 mg, 2.01 mmol), DMAP (8.08 mg, 0.066 mmol), and TEA (1.1 mL, 8.0 mmol) in MeCN (8 mL) and cooled to 0° C. was added TsCl (841 mg, 4.41 mmol) and the resulting mixture was stirred for 4 h. The reaction mixture was filtered and concentrated to dryness. The crude material was triturated with diethyl ether and the filtrate was collected and concentrated to dryness. The obtained product was purified via silica gel chromatography eluting with 0-80% EtOAc in heptane to afford 146-4 (670 mg, 1.51 mmol, 76% yield) as an off-white solid. MS [M+H]⁺=443.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.84-7.73 (m, 4H), 7.40-7.32 (m, 4H), 4.07 (t, J=6.2 Hz, 4H), 3.35 (t, J=6.0 Hz, 4H), 2.45 (s, 6H), 1.82 (quint, J=6.1 Hz, 4H).

Step 4. 3-(5-(((1S,2S)-2-(1,5-oxazocan-5-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-167)

To a solution of oxybis(propane-3,1-diyl) bis(4-methylbenzenesulfonate) (146-4, 193 mg, 0.437 mmol) and 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 100 mg, 0.291 mmol) in MeCN (2.5 mL) was added DIPEA (0.25 mL, 1.5 mmol) and the resulting solution was stirred at 120° C. under μW irradiation for 3 h. The reaction mixture was then concentrated to dryness and the obtained residue taken up in DCM and washed with saturated NaHCO₃ (aq). The organic phases were separated and the aqueous phase was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-60% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) and the obtained product was taken up in DCM and washed with sat. NaHCO₃ (aq) and the phases were separated via phase separator. The aqueous layer was extracted with DCM and the combined organic phases were concentrated to dryness. The crude material was dissolved in a minimal amount of EtOAc, cooled to 0° C. and diethyl ether was slowly added. The solvent was decanted and the solid material was dried under vacuum. The solid was purified further by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 10 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 25-50% MeCN, 3.5 min gradient; collected fractions contain ˜3 drops of formic acid) to afford the formate salt of I-167 (6.0 mg, 0.012 mmol, 4% yield) as a white solid. MS [M+H]⁺=442.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.94 (br s, 1H), 8.24 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.29-7.18 (m, 1H), 7.08-7.00 (m, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.71-4.63 (m, 1H), 4.43-4.19 (m, 2H), 3.63 (t, J=5.3 Hz, 4H), 2.96-2.84 (m, 1H), 2.68 (t, J=6.0 Hz, 4H), 2.64-2.55 (m, 1H), 2.43-2.35 (m, 2H), 2.16-2.03 (m, 1H), 2.03-1.85 (m, 2H), 1.75-1.55 (m, 7H), 1.55-1.44 (m, 1H).

Example 147: 3-(5-(((1S,2S)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1R,2R)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-168 and I-169)

Step 1. A mixture of 5-(((1S,2R)-2-hydroxycyclopentyl)oxy)isobenzofuran-1(3H)-one and 5-(((1R,2S)-2-hydroxycyclopentyl)oxy)isobenzofuran-1 (3H)-one (147-2)

To a mixture of 5-bromoisobenzofuran-1(3H)-one (19-1a, 1.00 g, 4.69 mmol), dtbbpy (63 mg, 0.24 mmol), NiCl₂(glyme) (52 mg, 0.24 mmol) and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (53 mg, 47 μmol) in a flask was added MeCN (15.7 mL) and the flask was evacuated and backfilled with nitrogen three times. Cis-1,2-pentanediol (147-1, 0.47 mL, 5.6 mmol) and 2,2,6,6-tetramethylpiperidine (0.83 mL, 4.9 mmol) were added and the flask was again evacuated and backfilled with nitrogen twice. The resulting mixture was stirred under blue LED light for 16 h, filtered, and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptanes to afford 147-2 (787 mg, 3.36 mmol, 72% yield, mixture of isomers) as a yellow solid. MS [M+H]⁺=235.2. ¹H NMR (400 MHz, Chloroform-d₆) δ 7.84 (d, J=8.5 Hz, 1H), 7.09 (dd, J=8.5, 2.2 Hz, 1H), 6.98 (d, J=2.1 Hz, 1H), 5.26 (s, 2H), 4.66 (dt, J=6.3, 4.6 Hz, 1H), 4.32 (td, J=5.7, 4.3 Hz, 1H), 2.16-2.06 (m, 1H), 2.04-1.79 (m, 4H), 1.74-1.61 (m, 1H).

Step 2. A mixture of (1S,2R)-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-methylbenzenesulfonate and (1R,2S)-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-methylbenzenesulfonate (147-3)

To a mixture of 5-(((1S,2R)-2-hydroxycyclopentyl)oxy)isobenzofuran-1(3H)-one and 5-(((1R,2S)-2-hydroxycyclopentyl)oxy)isobenzofuran-1(3H)-one (147-2, 0.100 g, 0.427 mmol) in DCM (3 mL) was added TsCl (179 mg, 0.939 mmol) and DIPEA (0.30 mL, 1.7 mmol) and the resulting mixture was stirred for 2 h at rt. 1-methylimidazole (34 μL, 0.43 mmol) was then added and the reaction mixture was stirred at rt for 2.5 h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate and extracted with dichloromethane (×3). The organic phases were combined, passed through a phase separator, and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptanes to afford 147-3 (787 mg, 3.36 mmol, 92% yield) as an orange solid. MS [M+H]⁺=389.3. ¹H NMR (400 MHz, Chloroform-d) δ 7.74-7.61 (m, 3H), 7.15 (d, J=8.1 Hz, 2H), 6.85 (dd, J=8.5, 2.2 Hz, 1H), 6.76 (d, J=2.1 Hz, 1H), 5.23-5.10 (m, 2H), 4.91 (td, J=6.1, 4.2 Hz, 1H), 4.63 (dt, J=5.9, 4.5 Hz, 1H), 2.33 (s, 3H), 2.15-1.87 (m, 5H), 1.70-1.58 (m, 1H).

Step 3. 5-(((1S,2S)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)isobenzofuran-1(3H)-one and 5-(((1R,2R)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)isobenzofuran-1(3H)-one (147-4 and 147-5)

To a μW vial with MeCN (0.3 mL) was added a mixture of (1S,2R)-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-methylbenzenesulfonate and (1R,2S)-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-methylbenzenesulfonate (147-3, 0.100 g, 0.257 mmol) and 3,3-dimethylpiperidine (0.04 mL, 0.3 mmol) and the vial was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at 150° C. for 3 h under μW irradiation, then quenched with water and extracted with dichloromethane (×3). The organic phases were combined, passed through a phase separator and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptanes to afford 147-4 (53.0 mg, 0.161 mmol, 63% yield) as a yellow oil and a mixture of isomer products. MS [M+H]⁺=330.3. The mixture of isomers were separated via chiral SFC [Column 2.1×25.0 cm Chiralpak AD-H; CO₂ Co-solvent MeOH and 5% Water with 0.25% Isopropylamine; Isocratic Method 20% Co-solvent at 80 g/min at 100 bar at 25° C.] to afford two enantiomers of trans isomer products: Peak 1: Enantiomer 1 of 2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-methylbenzenesulfonate (147-4, 15.0 mg, 0.045 mmol, 18% yield) as a yellow oil. Chiral SFC Rt 2.61 min. Peak 2: Enantiomer 2 of 2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-methylbenzenesulfonate (147-5, 18.0 mg, 0.054 mmol, 21% yield) as a yellow oil. Chiral SFC Rt 3.00 min. Absolute stereochemistry not determined.

Step 4. Single Enantiomer of Ethyl 2-(chloromethyl)-4-((˜2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)benzoate (147-6)

To a solution of a single enantiomer of 2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-methylbenzenesulfonate (Peak 1, 147-4, 15 mg, 0.045 mmol) in dichloroethane (1 mL) and EtOH (1 mL) stirred at 70° C. was added thionyl chloride (0.04 mL, 0.5 mmol) dropwise and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was cooled to rt, diluted with water and quenched with saturated aqueous sodium bicarbonate. The resulting mixture was extracted with EtOAc (×3) and the combined organic phases were passed through a phase separator and concentrated to dryness to afford 147-6 as a crude brown oil. The product was carried onto the next step without purification. MS [M+H]⁺=394.2.

Step 5. Single Enantiomer of 3-(5-((˜2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-168)

To a flask containing 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 15 mg, 0.090 mmol) in DMF (0.75 mL) was added DIPEA (0.04 mL, 0.2 mmol) and the flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at rt for 15 minutes, 147-6 (18 mg, 0.045 mmol) dissolved in DMF (1.1 mL) was added and the flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at 85° C. overnight and then stirred at 150° C. for 6 h under μW irradiation. The reaction mixture was concentrated to dryness and the crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane and then was further purified by reverse phase HPLC (Method Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 45-70% MeCN, 3.5 min gradient; collected fractions contain ˜3 drops of formic acid). The fractions containing the desired product were combined and lyophilized to afford I-168 (enantiomer 1, 2.1 mg, 4.2 μmol, 9% yield) as a white solid. MS [M+H]⁺=440.3. ¹H NMR (400 MHz, MeCN-d₃) δ 8.83 (s, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.22 (d, J=2.2 Hz, 1H), 7.08 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.4, 5.2 Hz, 1H), 4.78 (ddd, J=7.1, 4.2, 2.0 Hz, 1H), 4.34 (qd, J=16.7, 4.4 Hz, 2H), 3.01 (td, J=8.1, 7.6, 4.0 Hz, 1H), 2.84 (ddd, J=17.7, 13.3, 5.3 Hz, 2H), 2.74 (ddd, J=17.7, 4.8, 2.5 Hz, 1H), 2.56-2.36 (m, 3H), 2.30-2.19 (m, 2H), 2.17-2.00 (m, 2H), 1.82-1.57 (m, 6H), 1.31-1.24 (m, 2H), 0.96 (d, J=8.1 Hz, 6H).

Step 6. Single Enantiomer of Ethyl 2-(chloromethyl)-4-((-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)benzoate (147-7)

To a solution of a single enantiomer of 2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-methylbenzenesulfonate (Peak 2, 147-5, 18 mg, 0.054 mmol) in dichloroethane (1 mL) and EtOH (1 mL) stirred at 70° C. was added thionyl chloride (0.05 mL, 0.7 mmol) dropwise and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was cooled to rt, diluted with water, and quenched with saturated aqueous sodium bicarbonate. The resulting mixture was extracted with EtOAc (×3) and the combined organic phases were passed through a phase separator and concentrated to dryness to afford 147-7 as a crude brown oil. The product was carried onto the next step without purification. MS [M+H]⁺=394.4.

Step 7. Single Enantiomer of 3-(5-((-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-169)

To a flask containing 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 18 mg, 0.11 mmol) in DMF (0.75 mL) was added DIPEA (0.05 mL, 0.3 mmol) and the flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at rt for 15 minutes, then 147-7 (18 mg, 0.054 mmol) dissolved in DMF (1.1 mL) was added and the flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at 85° C. overnight then stirred at 150° C. for 6 h under μW irradiation. The reaction mixture was concentrated to dryness and the crude material purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane and then was further purified by reverse phase HPLC (Method Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 45-70% MeCN, 3.5 min gradient; collected fractions contain ˜3 drops of formic acid). The fractions containing the desired product were combined and lyophilized to afford I-169 (enantiomer 2, 1.5 mg, 2.9 μmol, 5% yield) as a white solid. MS [M+H]⁺=440.5. ¹H NMR (400 MHz, MeCN-d₃) δ 8.82 (s, 1H), 7.65 (d, J=8.4 Hz, 1H), 7.23 (d, J=2.2 Hz, 1H), 7.08 (dd, J=8.4, 2.2 Hz, 1H), 5.07 (dd, J=13.4, 5.2 Hz, 1H), 4.79-4.69 (m, 1H), 4.34 (qd, J=16.7, 4.5 Hz, 2H), 3.01-2.93 (m, 1H), 2.84 (ddd, J=17.7, 13.3, 5.4 Hz, 1H), 2.74 (ddd, J=17.7, 4.8, 2.5 Hz, 1H), 2.49-2.36 (m, 3H), 2.24-2.10 (m, 3H), 2.09-2.00 (m, 1H), 1.94-1.88 (m, 1H), 1.82-1.57 (m, 6H), 1.29-1.19 (m, 2H), 0.95 (d, J=8.7 Hz, 6H).

Example 148: 3-(5-(((1S,2S)-2-(ethyl(((1r,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-170)

To a solution of 3-(5-(((1S,2S)-2-((((1r,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl) oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-159, 0.10 g, 0.21 mmol) and acetaldehyde (2-1, 0.020 g, 0.43 mmol) in TFE (3 mL) was added NaBH(OAc)₃ (0.067 g, 0.32 mmol) at 0° C. and the resulting mixture was stirred at rt for 4 h. The reaction mixture was quenched with water and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 5% MeOH in DCM to afford I-170 (0.023 mg, 0.046 mmol, 23% yield) as an off-white solid. MS [M+H]⁺=498.4. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.60 (d, J=8.8 Hz, 1H), 7.24 (s, 1H), 7.07 (dd, J=8.8, 2.0 Hz, 1H), 5.08-5.06 (m, 1H), 4.62-4.59 (m, 1H), 4.40-.21 (m, 2H), 3.35-3.33 (m, 1H), 3.02 (s, 3H), 2.99 (m, 3H), 2.67-2.35 (m, 4H), 2.19-2.13 (m, 2H), 2.02-1.95 (m, 3H), 1.85-1.82 (m, 2H), 1.75-1.25 (m, 7H), 1.05-0.92 (m, 4H), 0.85-0.65 (m, 2H).

Example 149: 3-(5-(((1S,2S)-2-((((1R,4S)-4-methoxycyclohexyl)methyl)(methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-171)

To a solution of compound 3-(5-(((1S,2S)-2-((((1R,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-159, 0.10 g, 0.21 mmol) and formaldehyde (37% in water) (0.04 mL, 0.4 mmol) in TFE (4 mL) was added NaBH(OAc)₃ (0.067 g, 0.32 mmol) at 0° C. and the resulting mixture was stirred at rt for 16 h. The reaction mixture was concentrated to dryness and the residue was diluted with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified by silica gel chromatography eluting with 4% MeOH in DCM to afford I-171 (0.040 mg, 0.082 mmol, 39% yield) as an off-white solid. MS [M+H]⁺=484.3. ¹H NMR (400 MHz, DMSO-d₆): δ 10.98 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.21 (s, 1H), 7.07 (d, J=8.0 Hz, 1H), 5.09-5.04 (m, 1H), 4.68-4.65 (m, 1H), 4.41-4.23 (m, 2H), 3.19 (s, 3H), 3.0-2.67 (m, 3H), 2.67-2.33 (m, 2H), 2.15-2.08 (m, 5H), 2.07-1.96 (m, 4H), 1.83-1.36 (m, 6H), 1.51-1.48 (m, 2H), 1.25-1.22 (m, 2H), 0.83-0.80 (m, 2H).

Example 150: Mixture of 3-(5-(((1S,2S)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1R,2R)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-172)

Step 1. Zinc(II) Tetrahydroborate (150-2)

To a 100 mL round bottomed flask with a stir bar was added zinc chloride (150-1, 3.04 g, 22.3 mmol) and sodium borohydride (1.95 g, 51.4 mmol) and the flask was purged with nitrogen gas. THF (50 mL) was then added and the resulting mixture was stirred at rt for 3 days. The reaction mixture was filtered to afford 150-2 as a colorless solution (˜0.4 M) that was directly used in the next step without purification.

Step 2. A mixture of Methyl (1R,2R)-2-hydroxy-4,4-dimethylcyclopentane-1-carboxylate and methyl (1S,2S)-2-hydroxy-4,4-dimethylcyclopentane-1-carboxylate (150-4)

To methyl 4,4-dimethyl-2-oxocyclopentane-1-carboxylate (150-3, 420 mg, 2.47 mmol) dissolved in Et₂O (8 mL) and cooled to 0° C. was added zinc(II) tetrahydroborate (150-2, 7.5 mL, 3.0 mmol, 0.4M in THF) and the resulting solution was stirred at 0° C. for 1 hour. The reaction mixture was quenched with saturated aqueous ammonium acetate solution (10 mL) dropwise and then stirred for 15 minutes. The aqueous phase was extracted with dichloromethane, and the combined organic phases were passed through a phase separator and concentrated to dryness. The crude material purified via silica gel chromatography eluting with 0-40% EtOAc in heptane to afford 150-4 (284 mg, 1.65 mmol, 67% yield) as a mixture of trans isomers. MS [M+H]⁺=173.3. ¹H NMR (400 MHz, Chloroform-d) δ 4.45 (qd, J=7.9, 0.5 Hz, 1H), 3.72 (s, 3H), 2.83 (ddd, J=10.5, 8.9, 7.9 Hz, 1H), 1.97-1.79 (m, 2H), 1.73-1.63 (m, 1H), 1.55-1.44 (m, 1H), 1.11 (s, 3H), 1.04 (s, 3H).

Step 3. A mixture of Methyl (1R,2R)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carboxylate and methyl (1S,2S)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carboxylate (150-5)

To a mixture of methyl (1R,2R)-2-hydroxy-4,4-dimethylcyclopentane-1-carboxylate and methyl (1S,2S)-2-hydroxy-4,4-dimethylcyclopentane-1-carboxylate (150-4, 284 mg, 1.65 mmol) and imidazole (289 mg, 4.25 mmol) dissolved in DCM (6 mL) was added TBDMSCl (295 mg, 1.96 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was diluted with EtOAc (80 mL) and washed with water (20 mL), saturated aqueous sodium bicarbonate solution (20 mL) and brine (15 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated to afford 150-5 (522 mg, 1.73 mmol) which was carried onto the next step without purification. MS [M+H]⁺=287.3. ¹H NMR (400 MHz, DCM-d₂) δ 4.52-4.38 (m, 1H), 3.64 (s, 3H), 2.81 (ddd, J=10.0, 8.7, 6.9 Hz, 1H), 1.89-1.72 (m, 2H), 1.59 (dd, J=12.9, 10.1 Hz, 1H), 1.45 (ddd, J=12.9, 6.7, 1.0 Hz, 1H), 1.08 (s, 3H), 1.03 (s, 3H), 0.86 (s, 9H), 0.02 (d, J=3.5 Hz, 6H).

Step 4. A mixture of (1R,2R)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carboxylic Acid and (1S,2S)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carboxylic Acid (150-6)

A mixture of methyl (1R,2R)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carboxylate and methyl (1S,2S)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carboxylate (150-5, 522 mg, 1.82 mmol) and lithium hydroxide monohydrate (777 mg, 18.5 mmol) were dissolved in THF (3 mL), MeOH (3 mL), and Water (1 mL) and stirred at rt overnight. The reaction solution was diluted with water (60 mL) and acidified with 1 N HCl (aq) solution to a ˜pH 5. The aqueous mixture was extracted with EtOAc (3×100 mL) and the combined organic phases were dried over magnesium sulfate, filtered, and concentrated to afford 150-6 (465 mg, 1.62 mmol, 89% yield) as a colorless oil, which was carried onto the next step without purification. MS [M+H]⁺=273.4. ¹H NMR (400 MHz, DMSO-d₆) δ 12.05 (s, 1H), 4.43 (q, J=6.7 Hz, 1H), 2.66 (ddd, J=9.7, 8.9, 6.5 Hz, 1H), 1.83-1.69 (m, 2H), 1.49 (dd, J=12.8, 9.7 Hz, 1H), 1.37 (ddd, J=12.7, 6.5, 1.0 Hz, 1H), 1.05 (s, 3H), 0.99 (s, 3H), 0.84 (s, 9H), 0.01 (d, J=2.0 Hz, 6H).

Step 5. A mixture of tert-butyl (((1R,2R)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carbonyl)oxy)carbamate and tert-butyl (((1S,2S)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carbonyl)oxy)carbamate (150-8)

To a mixture of (1R,2R)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carboxylic acid and (1S,2S)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carboxylic acid (150-6, 465 mg, 1.71 mmol), N-Boc-hydroxylamine (150-7, 258 mg, 1.94 mmol), and HATU (692 mg, 1.82 mmol) dissolved in DCM (8 mL) was added DIPEA (0.75 mL, 4.3 mmol) was added and the resulting mixture was stirred at rt overnight. The reaction mixture was diluted with EtOAc (150 mL) and washed with water (30 mL), saturated aqueous sodium bicarbonate (30 mL), and brine (20 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-20% EtOAc in heptane to afford 150-8 (347 mg, 0.851 mmol, 50% yield) as a colorless oil. MS [M+H]⁺=386.5. ¹H NMR (400 MHz, DCM-d₂) δ 7.84 (s, 1H), 4.55 (dt, J=7.3, 6.4 Hz, 1H), 2.97 (ddd, J=9.8, 9.0, 6.6 Hz, 1H), 1.91 (ddd, J=13.0, 9.0, 1.1 Hz, 1H), 1.82 (dd, J=12.9, 7.3 Hz, 1H), 1.66 (dd, J=13.0, 9.8 Hz, 1H), 1.54-1.49 (m, 1H), 1.47 (s, 9H), 1.11 (s, 3H), 1.04 (s, 3H), 0.86 (s, 9H), 0.03 (d, J=2.1 Hz, 6H).

Step 6. A mixture of tert-butyl ((1R,2R)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentyl)carbamate and tert-butyl ((1S,2S)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentyl)carbamate (150-9)

A mixture of tert-butyl (((1R,2R)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carbonyl)oxy)carbamate and tert-butyl (((1S,2S)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentane-1-carbonyl)oxy)carbamate (150-8, 347 mg, 0.896 mmol) and cesium carbonate (336 mg, 1.03 mmol) in MeCN (7 mL) was stirred at 100° C. for 5 hours. The reaction mixture was then diluted with EtOAc and filtered through Celite®. The filtrate was concentrated to dryness and the crude material purified via silica gel chromatography eluting with 0-20% EtOAc in heptane to afford 150-9 (108 mg, 0.315 mmol, 35% yield) as a white solid. MS [M+H]⁺=344.4. ¹H NMR (400 MHz, DCM-d₂) δ 4.45 (s, 1H), 3.85-3.68 (m, 1H), 1.89 (dd, J=13.1, 7.8 Hz, 1H), 1.75 (dd, J=13.1, 7.2 Hz, 1H), 1.41 (s, 9H), 1.28-1.21 (m, 2H), 1.08 (s, 3H), 1.02 (s, 3H), 0.88 (s, 9H), 0.05 (d, J=4.9 Hz, 6H).

Step 7. A mixture of tert-butyl ((1R,2R)-2-hydroxy-4,4-dimethylcyclopentyl)carbamate and tert-butyl ((1S,2S)-2-hydroxy-4,4-dimethylcyclopentyl)carbamate (150-10)

To a mixture of tert-butyl ((1R,2R)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentyl)carbamate and tert-butyl ((1S,2S)-2-((tert-butyldimethylsilyl)oxy)-4,4-dimethylcyclopentyl)carbamate (150-9, 30.9 mg, 0.128 mmol) in THF (2 mL) was added TBAF (1M in THF) (0.350 mL, 0.350 mmol) and the resulting mixture was stirred at room temperature overnight. The reaction solution was concentrated to dryness and the crude material purified via silica gel chromatography eluting with 0-50% EtOAc in heptane to afford 150-10 (30.9 mg, 0.128 mmol, 52% yield). MS [M+H-t-Bu]⁺=174. ¹H NMR (400 MHz, Chloroform-d) δ 4.67 (s, 1H), 3.84-3.72 (m, 1H), 1.91 (ddd, J=11.6, 8.1, 3.5 Hz, 2H), 1.53 (dd, J=13.4, 7.4 Hz, 2H), 1.45 (s, 9H), 1.11 (s, 3H), 1.05 (s, 3H).

Step 8. A mixture of tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl) piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)-4,4-dimethylcyclopentyl)carbamate and tert-butyl ((1R,2R)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)-4,4-dimethylcyclopentyl)carbamate (150-11)

To a mixture of tert-butyl ((1R,2R)-2-hydroxy-4,4-dimethylcyclopentyl)carbamate and tert-butyl ((1S,2S)-2-hydroxy-4,4-dimethylcyclopentyl)carbamate (150-10, 30.0 mg, 0.131 mmol), 3-(5-bromo-1-oxoisoindolin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (31-3a, 62.0 mg, 0.137 mmol), NiCl₂(glyme) (1.4 mg, 6.5 μmol), dtbbpy (1.8 mg, 6.5 μmol), and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (1.5 mg, 1.3 μmol) purged with nitrogen was added MeCN (1 mL) and 2,2,6,6-tetramethylpiperidine (40-2, 0.024 mL, 0.14 mmol) and the resulting mixture was stirred under blue LED lights at rt for 18 hours. The reaction mixture was diluted with EtOAc and filtered through Celite®, rinsing with EtOAc. The filtrate was concentrated to dryness and purified via silica gel chromatography eluting with 0-100% EtOAc in heptane to afford 150-11 (29.1 mg, 0.034 mmol, 26% yield) as a yellow solid. MS [M+H₂O]⁺=619.5. ¹H NMR (400 MHz, DCM-d₂) δ 7.71 (dd, J=8.6, 4.2 Hz, 1H), 7.02 (d, J=6.8 Hz, 2H), 5.25-5.05 (m, 2H), 4.66 (s, 1H), 4.38-4.30 (m, 2H), 3.69-3.50 (m, 2H), 3.06-2.77 (m, 2H), 2.31 (dd, J=13.7, 4.7 Hz, 1H), 2.21-2.13 (m, 1H), 2.11-2.01 (m, 2H), 1.66 (dd, J=13.9, 4.2 Hz, 1H), 1.48-1.44 (m, 3H), 1.41 (s, 9H), 1.13 (d, J=7.9 Hz, 6H), 0.95-0.89 (m, 2H), 0.00 (s, 9H).

Step 9. A mixture of 3-(5-(((1S,2S)-2-amino-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1R,2R)-2-amino-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (150-12)

To a mixture of tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl) piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)-4,4-dimethylcyclopentyl)carbamate and tert-butyl ((1R,2R)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)-4,4-dimethylcyclopentyl)carbamate (150-11, 29 mg, 0.048 mmol) in MeCN (1 mL) was added methanesulfonic acid (0.013 mL, 0.20 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was cooled to 0° C. and whilst under a stream of nitrogen, TEA (0.067 mL, 0.48 mmol) was added. The solution was warmed to room temperature and then stirred for 30 minutes. N1,N2-dimethylethane-1,2-diamine (6 μL, 0.05 mmol) was then added and the resulting mixture was stirred at rt overnight. The reaction mixture was concentrated to approximately half volume and a 1:1 mixture of saturated aqueous sodium bicarbonate and water (30 mL) was added. The aqueous mixture was extracted with DCM/isopropanol 4:1 mixture (3×30 mL) and the combined organic phases were dried over magnesium sulfate, filtered, and concentrated to dryness to afford 150-12 as a yellow solid, which was carried onto the next step without purification. MS [M+H]⁺=372.4.

Step 10. A mixture of 3-(5-(((1S,2S)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1R,2R)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-172)

To a mixture of 3-(5-(((1S,2S)-2-amino-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1R,2R)-2-amino-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (150-12, 18 mg, 0.048 mmol) and acetaldehyde (2-1, 8 μL, 0.15 mmol) dissolved in DMF (0.5 mL) was added sodium triacetoxyborohydride (56 mg, 0.26 mmol) in one portion and the resulting mixture was stirred at rt overnight. The reaction mixture was then diluted with EtOAc (45 mL) and washed with saturated aqueous sodium bicarbonate solution (10 mL) and brine (10 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0 to 80% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) and then further purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford the formate salt of I-172 (6.0 mg, 0.012 mmol, 25% yield) as a white solid. MS [M+H]⁺=428.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 8.18 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.16 (s, 1H), 7.02 (d, J=8.4 Hz, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.76-4.63 (m, 1H), 4.45-4.18 (m, 2H), 3.48 (dt, J=11.8, 6.3 Hz, 2H), 2.97-2.83 (m, 1H), 2.63-2.51 (m, 3H), 2.44-2.31 (m, 2H), 1.98 (dd, J=14.3, 7.5 Hz, 2H), 1.72 (dd, J=12.3, 7.2 Hz, 1H), 1.55-1.35 (m, 2H), 1.06 (d, J=4.6 Hz, 6H), 0.94 (t, J=7.0 Hz, 6H).

Example 151: 3-(5-(((1S,2S)-2-(((4,4-difluorocyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-173)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 100 mg, 0.291 mmol) in MeCN (1.5 mL) was added 4-(bromomethyl)-1,1-difluorocyclohexane (151-1, 68 mg, 0.32 mmol) and DIPEA (153 μL, 0.874 mmol) and the resulting mixture was stirred at 140° C. for 45 minutes in the μW. NaI (48 mg, 0.32 mmol) was added and stirring was continued at 140° C. for 2 h and 45 minutes in the μW. The reaction mixture was then added to a saturated solution of aqueous sodium hydrogen carbonate (20 mL) and extracted with 20% i-PrOH in DCM (×2). The combined organic phases were passed through a phase separating column and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-173 (20 mg, 0.040 mmol, 14% yield) as a cream-colored solid. MS [M+H]⁺=476.4. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.17 (s, 1H), 7.04 (d, J=8.4 Hz, 1H), 5.11-4.97 (m, 1H), 4.65-4.52 (m, 1H), 4.44-4.19 (m, 2H), 3.18-3.03 (m, 1H), 2.90 (ddd, J=18.3, 13.6, 5.4 Hz, 1H), 2.59 (d, J=17.6 Hz, 1H), 2.47-2.28 (m, 2H), 2.17-2.05 (m, 1H), 2.04-1.87 (d, J=13.0 Hz, 4H), 1.86-1.58 (m, 8H), 1.56-1.38 (m, 2H), 1.21-1.07 (m, 2H).

Example 152: 3-(5-(((1S,2S)-2-(((1H-indol-5-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-174)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 100 mg, 0.291 mmol) in DMF (2 mL) under an atmosphere of nitrogen, was added 1H-indole-5-carbaldehyde (152-1, 46.5 mg, 0.320 mmol) and the resulting mixture was stirred at rt for 30 minutes. Sodium triacetoxyborohydride (185 mg, 0.874 mmol) was added and stirring was continued at rt overnight. The reaction mixture was then added to a saturated aqueous solution of sodium hydrogen carbonate (30 mL) and extracted with 20% i-PrOH in DCM (×3). The combined organic phases were passed through a phase separating column and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-20% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) and then further purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-174 (12 mg, 0.023 mmol, 8% yield) as a cream-colored solid. MS [M+H]⁺=473.4. ¹H NMR (400 MHz, DMSO-d₆) δ (ppm): 10.95 (s, 2H), 7.56 (dd, J=8.4, 1.9 Hz, 1H), 7.48 (s, 1H), 7.31 (d, J=11.9 Hz, 1H), 7.28 (s, 1H), 7.11-7.05 (m, 2H), 6.99 (dd, J=8.4, 2.4 Hz, 1H), 6.37-6.33 (m, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.69-4.62 (m, 1H), 4.36-4.09 (m, 2H), 3.95-3.68 (m, 2H), 3.20-3.12 (m, 1H), 2.91 (ddd, J=18.1, 13.7, 5.4 Hz, 1H), 2.60 (d, J=17.1 Hz, 1H), 2.39 (td, J=13.1, 4.5 Hz, 1H), 2.21-2.09 (m, 1H), 2.04-1.84 (m, 2H), 1.84-1.60 (m, 3H), 1.53 (d, J=9.3 Hz, 1H).

Example 153: 3-(5-(((1S,2S)-2-(4-(tert-butoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-175)

Tert-butyl 2,2,2-trichloroacetimidate (153-1, 5.0 μL, 0.028 mmol) was added to a suspension of 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-96, 10 mg, 0.023 mmol) and scandium triflate (1.2 mg, 2.3 μmol) in dry DCM (0.25 mL) and the resulting mixture was stirred at rt for 24 h. Further scandium triflate (1.2 mg, 2.3 μmol) and tert-butyl 2,2,2-trichloroacetimidate (5 μL, 0.03 mmol) were added and stirring was continued at rt for several days. The reaction mixture was then filtered through Celite® and concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-175 (3.0 mg, 5.3 μmol, 23% yield) as a white solid. MS [M+H]⁺=484.4. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 8.07 (s, 1H), 7.71 (d, J=8.3 Hz, 1H), 7.08-6.95 (m, 2H), 5.18-5.07 (m, 1H), 4.91 (s, 1H), 4.43-4.22 (m, 2H), 3.55 (s, 1H), 3.25-2.74 (m, 6H), 2.60-2.24 (m, 3H), 2.24-2.01 (m, 3H), 1.94-1.66 (m, 6H), 1.56 (s, 2H), 1.16 (s, 9H).

Example 154: Single Enantiomer of 3-(5-((2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-176)

Step 1. Single Enantiomer of 5-((2-hydroxycyclopentyl)oxy)isobenzofuran-1(3H)-one (154-1)

5-bromoisobenzofuran-1(3H)-one (19-1a, 2.50 g, 11.7 mmol), dtbbpy (157 mg, 0.587 mmol), NiCl₂(glyme) (129 mg, 0.587 mmol), and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (131 mg, 0.117 mmol) and MeCN (35 mL) were placed in a flask and the flask was then evacuated and backfilled with nitrogen three times. Cis-1,2-pentanediol (147-1, 1.17 mL, 14.1 mmol) and 2,2,6,6-tetramethylpiperidine (40-2, 2.1 mL, 12 mmol) were added and the flask was again evacuated and backfilled with nitrogen twice more. The resulting mixture was stirred under blue LED lights at rt for 18 h. The reaction mixture was filtered, concentrated onto Celite® and purified by silica gel chromatography eluting with 0-100% EtOAc in heptane to afford the desired product (1.96 g, 8.37 mmol, 71% yield) as a yellow solid as a mixture of cis isomers. MS [M+H]⁺=235.3. The mixture of isomers was separated via chiral SFC [Column: 2.1×25.0 cm Chiralpak AD-H; CO₂ Co-solvent: 20% MeOH: Flow rate 80 g/min at 125 bar at 25° C.] to afford two cis isomers: Peak 1: 154-1 (1.01 g, 4.32 mmol, 37% yield) as a pale yellow oil. Chiral SFC Rt=1.71 min. Peak 2: 154-2 (1.03 g, 4.38 mmol, 37% yield) as a pale yellow oil. Chiral SFC Rt=1.87 min. Absolute stereochemistry not determined.

Step 2. Single Enantiomer of 2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl 4-nitrobenzenesulfonate (154-4)

To a solution of 154-1 (Peak 1, 0.500 g, 2.13 mmol) in DCM (14.2 mL) was added 4-nitrobenzenesulfonyl chloride (154-3, 1.42 g, 6.40 mmol) and 1-methylimidazole (0.34 mL, 4.27 mmol) and the resulting mixture was stirred at rt for 20 h. Additional 4-nitrobenzenesulfonyl chloride (378 mg, 1.71 mmol) and 1-methylimidazole (0.17 mL, 2.1 mmol) were added and stirring was continued at rt for 2 h. The reaction mixture was then quenched with saturated aqueous sodium bicarbonate and extracted with dichloromethane (×3). The combined organic phases were passed through a phase separator and concentrated onto Celite®. The Celite® residue was purified by silica gel chromatography eluting with 0-100% EtOAc in heptane and was further purified by silica gel chromatography eluting with 0-100% EtOAc in heptane to afford 154-4 (738 mg, 1.76 mmol, 82% yield) as a white solid. MS [M+H]⁺=420.2.

Step 3. Single Enantiomer of 5-((2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)isobenzofuran-1(3H)-one (154-6)

A suspension of 2-oxa-8-azaspiro[4.5]decane hydrochloride (154-5, 83.0 mg, 0.465 mmol) and Cs₂CO₃ (0.186 g, 0.572 mmol) in MeCN (1.4 mL) was stirred at rt for 20 minutes. The suspension was then then added to 154-4 (0.150 g, 0.358 mmol) followed by the addition of DIPEA (0.094 mL, 0.54 mmol). The resulting mixture was purged with nitrogen with sonication for 20 minutes and then stirred at 120° C. for 3 h under μW radiation. The reaction mixture was quenched with water and extracted with dichloromethane. The combined organic phases were passed through a phase separator and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptane and then with 0-20% MeOH in DCM to afford 154-6 (81.5 mg, 0.228 mmol, 64% yield) as a cream-colored solid. MS [M+H]⁺=358.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.79 (d, J=8.5 Hz, 1H), 7.00 (dd, J=8.5, 2.1 Hz, 1H), 6.95-6.85 (m, 1H), 5.24 (s, 2H), 4.91-4.74 (m, 1H), 3.83 (t, J=7.1 Hz, 2H), 3.53 (s, 2H), 3.03-2.92 (m, 1H), 2.67-2.48 (m, 4H), 2.15-2.05 (m, 2H), 1.83-1.59 (m, 10H).

Step 4. Single Enantiomer of Ethyl 4-((2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-2-(chloromethyl)benzoate (154-7)

To a stirred suspension of 154-6 (81.5 mg, 0.228 mmol) in dichloroethane (1.1 mL) and EtOH (1.1 mL) at 70° C. was added thionyl chloride (0.20 mL, 2.7 mmol) dropwise and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was then cooled to rt, diluted with water, and quenched with saturated aqueous sodium bicarbonate. The aqueous layer was extracted with EtOAc (×3) and the combined organic phases were passed through a phase separator and concentrated to dryness to afford 154-7 as a crude brown oil. The crude product was carried on to the next step without purification. MS [M+H]⁺=422.4.

Step 5. Single Enantiomer of 3-(5-((2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-176)

To a flask containing 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 75.0 mg, 0.456 mmol) in DMF (0.9 mL) was added DIPEA (0.2 mL, 1 mmol) and the flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at rt for 15 minutes, 154-7 (96 mg, 0.23 mmol) dissolved in DMF (1.37 mL) was added and the flask was again evacuated and backfilled with nitrogen three times. The reaction mixture was stirred at 85° C. overnight and then at 150° C. for 5 h under μW irradiation. The reaction mixture was concentrated to dryness and purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane to afford I-176 (61.7 mg, 0.128 mmol, 56% yield) as a cream-colored solid. MS [M+H]⁺=468.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (d, J=2.2 Hz, 1H), 7.03 (dt, J=8.7, 1.9 Hz, 1H), 5.07 (dd, J=13.4, 5.1 Hz, 1H), 4.77-4.63 (m, 1H), 4.38 (dd, J=17.2, 9.6 Hz, 1H), 4.25 (dd, J=17.2, 7.9 Hz, 1H), 3.69 (t, J=7.1 Hz, 2H), 3.44 (dd, J=11.6, 5.1 Hz, 1H), 3.39 (s, 2H), 2.97-2.80 (m, 2H), 2.63-2.53 (m, 2H), 2.46-2.34 (m, 4H), 2.06-1.90 (m, 3H), 1.70-1.57 (m, 5H), 1.51-1.45 (m, 4H). Absolute stereochemistry not determined.

Example 155: 3-(5-(((1S,2S)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-177)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 50.0 mg, 0.146 mmol) and 2-(2-chlorophenoxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (94-3, 74.4 mg, 0.146 mmol) in MeCN (0.75 mL) was added DIPEA (0.10 mL, 0.57 mmol) and the resulting mixture was stirred at 120° C. under μW irradiation for 8 h and then concentrated. The crude product was purified via silica gel chromatography eluting with 0-50% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-177 (34.0 mg, 0.063 mmol, 44% yield) as a white solid. MS [M+H]⁺=510.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.34-7.26 (m, 2H), 7.14 (d, J=2.2 Hz, 1H), 7.03 (dd, J=8.3, 2.2 Hz, 1H), 6.89-6.82 (m, 2H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.77 (quint, J=5.5 Hz, 1H), 4.61-4.54 (m, 1H), 4.38 (dd, J=17.2, 5.7 Hz, 1H), 4.25 (dd, J=17.2, 4.9 Hz, 1H), 3.73 (q, J=7.9, 7.4 Hz, 2H), 3.13-3.01 (m, 2H), 2.98-2.82 (m, 2H), 2.65-2.54 (m, 1H), 2.43-2.35 (m, 1H), 2.20-2.07 (m, 1H), 2.05-1.90 (m, 1H), 1.85-1.75 (m, 1H), 1.75-1.55 (m, 3H), 1.50-1.36 (m, 1H).

Example 156: 3-(5-(((1S,2S)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-178)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 33 mg, 0.099 mmol) and 2-(2-methoxyphenoxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (95-3, 50 mg, 0.099 mmol) in MeCN (0.75 mL) was added DIPEA (0.052 mL, 0.30 mmol) and resulting mixture was stirred at 120° C. under μW irradiation for 8 h. The reaction mixture was concentrated to dryness and purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-178 (16 mg, 0.028 mmol, 29% yield) as a pale yellow solid. MS [M+H]⁺=506.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.15 (s, 1H), 7.04 (d, J=8.4 Hz, 1H), 6.97 (d, J=8.0 Hz, 1H), 6.91 (d, J=8.1 Hz, 1H), 6.86-6.79 (m, 1H), 6.73 (d, J=7.8 Hz, 1H), 5.10-5.01 (m, 1H), 4.75-4.66 (m, 1H), 4.62-4.54 (m, 1H), 4.39 (dd, J=17.2, 5.4 Hz, 1H), 4.26 (dd, J=17.3, 5.3 Hz, 1H), 3.74 (m, 5H), 3.15-3.00 (m, 2H), 2.90 (ddd, J=18.4, 13.4, 5.4 Hz, 2H), 2.63-2.54 (m, 1H), 2.46-2.33 (m, 1H), 2.21-2.07 (m, 1H), 2.04-1.92 (m, 1H), 1.84-1.54 (m, 4H), 1.49-1.38 (m, 1H).

Example 157: Single Enantiomer of 3-(5-((2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-179)

Step 1. Single Enantiomer of 5-((2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)isobenzofuran-1(3H)-one (157-2)

To a suspension of 154-4 (Example 154, Step 2, 0.150 g, 0.358 mmol) in MeCN (1.2 mL) was added sequentially 6-azaspiro[3.5]nonane (157-1, 54.0 mg, 0.429 mmol), and DIPEA (0.094 mL, 0.54 mmol) and the resulting mixture was purged with nitrogen with sonication for 20 minutes and then stirred at 120° C. for 3 h under μW radiation. The reaction mixture was quenched with water and extracted with dichloromethane. The combined organic phases were passed through a phase separator and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptane to afford 157-2 (83.0 mg, 0.243 mmol, 68% yield) as a pale brown solid. MS [M+H]⁺=341.9. ¹H NMR (400 MHz, Chloroform-d) δ 7.84-7.76 (m, 1H), 7.13-7.04 (m, 2H), 5.24 (s, 2H), 4.84-4.60 (m, 1H), 3.04-2.91 (m, 1H), 2.55-2.21 (m, 4H), 2.06-1.93 (m, 2H), 1.91-1.59 (m, 10H), 1.56-1.43 (m, 4H).

Step 2. Single Enantiomer of Ethyl 4-((2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)-2-(chloromethyl)benzoate (157-3)

A suspension of 157-2 (83.0 mg, 0.243 mmol) in dichloroethane (1.2 mL) and Ethanol (1.2 mL) was stirred at 70° C. and thionyl chloride (0.21 mL, 2.9 mmol) was then added dropwise. The resulting mixture was stirred at 70° C. overnight and then cooled to rt, diluted with water. and quenched with saturated aqueous sodium bicarbonate. The reaction mixture was extracted with EtOAc (×3). The combined organic phases were passed through a phase separator and concentrated to dryness to afford 157-3 as a crude brown oil. The crude product was taken on to the next step without purification. MS [M+H]⁺=406.3.

Step 3. Single Enantiomer of cis isomer of 3-(5-((2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-179)

To a flask containing a solution of 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 80.0 mg, 0.486 mmol) in DMF (1 mL) was added DIPEA (0.2 mL, 1 mmol) and the flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at rt for 15 minutes and then 157-3 (99.0 mg, 0.243 mmol) dissolved in DMF (1.5 mL) was added and the flask was again evacuated and backfilled with nitrogen three times. The reaction mixture was stirred at 85° C. overnight and then stirred at 150° C. for 4 h under μW irradiation. The reaction mixture was concentrated to dryness and the crude material purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in heptane (with 0.1% NEt₃) to afford crude material. The material was further purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane to afford I-179 (21.8 mg, 0.045 mmol, 19% yield) as a white solid. MS [M+H]⁺=452.5. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.60 (d, J=8.5 Hz, 1H), 7.26 (d, J=2.2 Hz, 1H), 7.07 (dt, J=8.5, 2.6 Hz, 1H), 5.07 (ddd, J=13.4, 5.2, 1.9 Hz, 1H), 4.77-4.64 (m, 1H), 4.38 (dd, J=17.1, 9.1 Hz, 1H), 4.25 (dd, J=17.2, 6.7 Hz, 1H), 2.96-2.84 (m, 2H), 2.63-2.55 (m, 1H), 2.43-2.23 (m, 5H), 2.07-1.92 (m, 2H), 1.92-1.74 (m, 3H), 1.72-1.57 (m, 7H), 1.56-1.46 (m, 1H), 1.45-1.35 (m, 4H). Absolute stereochemistry not determined

Example 158: 3-(5-(((1S,2S)-2-(ethyl(((1s,3R)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-180)

Step 1. ((1S,3S)-3-methoxycyclobutyl)methanol (158-2)

To a solution of cis-3-methoxycyclobutane-1-carboxylic acid (158-1, 1.0 g, 7.7 mmol) in THF (20 mL) at 0° C. was added 1M BH₃ in THF (17 mL, 17 mmol). The resulting mixture was allowed to warm slowly to rt and then stirred at rt for 18 h. The reaction mixture was slowly quenched by adding MeOH dropwise and then stirred for 20 min. The mixture was then concentrated to dryness. The crude material was dissolved in MeOH (50 mL), stirred at rt for 3 h and then concentrated to dryness to afford crude 158-2 (910 mg, 7.8 mmol). The crude product was taken on to the next step without purification. ¹H NMR (400 MHz, DMSO-d₆) δ 4.45 (t, J=5.4 Hz, 1H), 3.76-3.56 (m, 1H), 3.35-3.29 (m, 2H), 3.08 (s, 3H), 2.22-2.06 (m, 2H), 1.97-1.77 (m, 1H), 1.51 (tdd, J=9.2, 7.7, 2.7 Hz, 2H).

Step 2. ((1S,3S)-3-methoxycyclobutyl)methyl 4-methylbenzenesulfonate (158-3)

To a solution of ((1S,3S)-3-methoxycyclobutyl)methanol (158-2, 910 mg, 7.83 mmol) in pyridine (12 mL) was added tosyl chloride (2.20 g, 11.8 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was diluted with EtOAc and washed with water. The phases were separated and the aqueous phase was extracted with EtOAc (×1). The combined organics were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc/heptanes to afford 158-3 (1.40 g, 5.18 mmol, 66% yield) as a colorless oil. MS [M+H]⁺=271.2. ¹H NMR (400 MHz, DMSO-d₆) δ 7.77 (d, J=8.4 Hz, 2H), 7.57-7.43 (m, 2H), 3.97 (d, J=6.1 Hz, 2H), 3.73-3.52 (m, 1H), 3.05 (s, 3H), 2.42 (s, 3H), 2.28-1.94 (m, 3H), 1.56-1.37 (m, 2H).

Step 3. 3-(5-(((1S,2S)-2-(ethyl(((1S,3R)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-180)

To 3-(5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-29, 91 mg, 0.25 mmol) and ((1S,3S)-3-methoxycyclobutyl)methyl 4-methylbenzenesulfonate (158-3, 199 mg, 0.735 mmol) dissolved in DMF (0.6 mL) was added DIPEA (0.2 mL, 1 mmol) and the resulting mixture was stirred at rt for 18 h and then stirred at 60° C. for 16 h. TBAI (catalytic amount) and further ((1S,3S)-3-methoxycyclobutyl)methyl 4-methylbenzenesulfonate (158-3, ˜150-200 mg) were added and stirring was continued at 100° C. for 3 days. Further ((1s,3s)-3-methoxycyclobutyl)methyl 4-methylbenzenesulfonate (158-3, −200 mg) and TBAI was added and stirring was continued at 100° C. for 3 h. Further DIPEA (200 μL) was added and stirring was continued at 100° C. for 4 h. The reaction mixture was cooled to rt, diluted with acetone, and concentrated to dryness onto Celite®. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1) in DCM and the further purified via reverse phase chromatography on C-18 column eluting with 10-60% MeCN in water with 0.1% formic acid as a modifier. The obtained material was purified further via reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 5-95% MeCN, 3.5 min gradient) to afford I-180 (22 mg, 0.045 mmol, 18% yield) as a white solid. MS [M+H]⁺=470.3. ¹H NMR (400 MHz, DMSO-d₆) δ 8.31 (s, 1H), 7.60 (d, J=8.3 Hz, 1H), 7.28 (d, J=1.9 Hz, 1H), 7.08 (dt, J=8.4, 2.4 Hz, 1H), 5.08 (ddd, J=13.6, 5.1, 2.1 Hz, 1H), 4.67-4.56 (m, 1H), 4.46-4.16 (m, 2H), 3.65 (t, J=7.2 Hz, 1H), 3.24 (tt, J=8.8, 4.0 Hz, 2H), 3.07 (s, 3H), 3.01-2.81 (m, 1H), 2.72-2.56 (m, 1H), 2.45-2.19 (m, 5H), 2.07-1.78 (m, 5H), 1.73-1.57 (m, 3H), 1.54-1.34 (m, 3H), 0.95 (t, J=7.0 Hz, 3H).

Example 159: Single Enantiomer of trans-3-(5-((2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-181)

Step 1. A mixture of tert-butyl ((1S,2S)-4,4-dimethyl-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate and tert-butyl ((1S,2S)-4,4-dimethyl-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate (159-1)

To 5-bromoisobenzofuran-1(3H)-one (19-1a, 43.2 mg, 0.203 mmol), a mixture of tert-butyl ((1R,2R)-2-hydroxy-4,4-dimethylcyclopentyl)carbamate and tert-butyl ((1S,2S)-2-hydroxy-4,4-dimethylcyclopentyl)carbamate (150-10, 42 mg, 0.18 mmol), NiCl₂(glyme) (2 mg, 9 μmol), dtbbpy (2 mg, 9 μmol), and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (2 mg, 2 μmol) under an atmosphere of nitrogen was added MeCN (1 mL) and 2,2,6,6-tetramethylpiperidine (40-2, 34 μL, 0.20 mmol) and the resulting mixture was stirred under blue LED lights at rt for 2 h. The reaction mixture was diluted with EtOAc and filtered through Celite®. The filtrate was collected and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-70% EtOAc in heptane to afford 159-1 (47.1 mg, 0.117 mmol, 64% yield) as a white solid. MS [M+H]⁺=362.4. ¹H NMR (400 MHz, DCM-d₂) δ 7.79 (d, J=8.5 Hz, 1H), 7.09 (d, J=8.6 Hz, 1H), 7.02 (s, 1H), 5.25 (s, 2H), 4.78-4.67 (m, 1H), 4.32-4.20 (m, 1H), 2.09 (dd, J=13.9, 7.2 Hz, 2H), 1.70 (dd, J=14.0, 4.1 Hz, 1H), 1.53-1.48 (m, 1H), 1.44 (s, 9H), 1.18 (s, 3H), 1.17 (s, 3H).

Step 2. A mixture of 5-(((1S,2S)-2-amino-4,4-dimethylcyclopentyl)oxy)isobenzofuran-1(3H)-one and 5-(((1S,2S)-2-amino-4,4-dimethylcyclopentyl)oxy)isobenzofuran-1 (3H)-one (159-2)

To a mixture of tert-butyl ((1S,2S)-4,4-dimethyl-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate and tert-butyl ((1S,2S)-4,4-dimethyl-2-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate (159-1, 47.1 mg, 0.130 mmol) in DCM (1 mL) was added TFA (0.100 mL, 1.30 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was concentrated to dryness and azeotroped with MeOH and DCM to afford 159-2 (77.8 mg, 0.176 mmol) as a crude solid. The product was taken on to the next step without purification. MS [M+H]⁺=262.3.

Step 3. 5-(((1S,2S)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)isobenzofuran-1(3H)-one and 5-(((1R,2R)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)isobenzofuran-1(3H)-one (159-3 and 159-4)

To a mixture of 5-(((1S,2S)-2-amino-4,4-dimethylcyclopentyl)oxy)isobenzofuran-1(3H)-one and 5-(((1S,2S)-2-amino-4,4-dimethylcyclopentyl)oxy)isobenzofuran-1(3H)-one (159-2, 49 mg, 0.13 mmol) and acetaldehyde (2-1, 0.022 mL, 0.39 mmol) dissolved in DMF (1 mL) was added sodium triacetoxyborohydride (0.14 g, 0.65 mmol) in one portion and the resulting mixture was stirred at rt overnight. The reaction mixture was diluted with EtOAc (45 mL) and washed with saturated aqueous sodium bicarbonate solution (10 mL) and brine (10 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 5-100% EtOAc (with 0.1% NEt₃) in heptane (with 0.1% NEt₃) to afford a mixture two trans isomers. The trans isomers were separated via chiral SFC (Method: Column Chiralpak IE (5 uM) 21×250 mm Mobile Phase: 40% MeOH w/10 mM NH₃/CO₂ 80g/min 125 bar 6.2 min; Injection 5.6 mg (11.2 mg/mL in MeOH) to afford two products: Peak 1: 159-3 (14.2 mg, 0.041 mmol) as a solid. Chiral SFC Rt 1.02 min. MS [M+H]⁺=318.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.82 (d, J=8.4 Hz, 1H), 7.12-7.03 (m, 1H), 6.99 (s, 1H), 5.66 (s, 1H), 5.28 (s, 2H), 3.87 (s, 1H), 3.40-3.23 (m, 2H), 3.05 (dt, J=13.5, 6.7 Hz, 2H), 2.44-2.27 (m, 2H), 1.98 (t, J=9.9 Hz, 1H), 1.72 (d, J=14.4 Hz, 1H), 1.37 (dd, J=8.3, 6.1 Hz, 6H), 1.22 (s, 3H), 1.16 (s, 3H). Peak 2: 159-4 (8.2 mg, 0.025 mmol) as a solid. Chiral SFC Rt 1.22 min. MS [M+H]⁺=318.3. ¹H NMR (400 MHz, DCM-d₂) δ 7.82 (d, J=8.5 Hz, 1H), 7.07 (dd, J=8.4, 2.2 Hz, 1H), 6.99 (s, 1H), 5.74-5.61 (m, 1H), 5.29 (d, J=1.7 Hz, 2H), 3.87 (s, 1H), 3.32 (dt, J=12.8, 6.6 Hz, 2H), 3.04 (dq, J=13.5, 6.6 Hz, 2H), 2.46-2.25 (m, 2H), 2.03-1.91 (m, 1H), 1.78-1.69 (m, 1H), 1.41-1.34 (m, 6H), 1.22 (s, 3H), 1.16 (s, 3H). Absolute stereochemistry not determined.

Step 4. Single Enantiomer of trans-Ethyl 2-(chloromethyl)-4-((2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)benzoate (159-5)

To 159-4 (Peak 2, 8.2 mg, 0.026 mmol) dissolved in Ethanol (1 mL) was added thionyl chloride (0.010 mL, 0.14 mmol) and the resulting mixture was stirred at 70° C. overnight. Additional thionyl chloride (0.010 mL, 0.14 mmol) was added and stirring was continued at 70° C. overnight. The reaction mixture was concentrated to dryness to afford 159-5. The crude material was taken on to the next step without purification. MS [M+H]⁺=382.3.

Step 5. Single Enantiomer of trans-3-(5-((2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-181)

To a solution of 159-5 (9.9 mg, 0.026 mmol) in DMF (0.5 mL) and DIPEA (0.030 mL, 0.17 mmol) under an atmosphere of nitrogen was added 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 6.4 mg, 0.039 mmol). The resulting mixture was sparged with nitrogen and stirred at 110° C. overnight. The reaction mixture was then diluted with EtOAc (40 mL) and washed with saturated aqueous sodium bicarbonate solution (2×10 mL) and brine (10 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid, 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) then further purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in DCM (with 1% NEt₃) to afford I-181 (2.3 mg, 4.6 μmol) as a white solid. MS [M+H]⁺=428.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.23-7.15 (m, 1H), 7.03 (dd, J=8.4, 2.0 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.74-4.63 (m, 1H), 4.46-4.16 (m, 2H), 3.56-3.42 (m, 1H), 1H obscured by asymmetric DMSO signal, 2.91 (ddd, J=18.2, 13.5, 5.4 Hz, 1H), 1.98 (dd, J=13.8, 8.4 Hz, 2H), 1.72 (dd, J=12.4, 7.3 Hz, 1H), 1.58-1.38 (m, 2H), 1.06 (d, J=4.6 Hz, 6H), 0.95 (t, J=7.0 Hz, 6H). Absolute stereochemistry not determined

Example 160: 3-(5-(((1S,2S)-2-(ethyl(((1r,3S)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-182)

Step 1. ((1r,3r)-3-methoxycyclobutyl)methyl 4-methylbenzenesulfonate (160-1)

To a solution of (3-methoxycyclobutyl)methanol (81-2, 1.40 g, 7.83 mmol) in pyridine (13 mL) was added TsCl (2.20 g, 12.0 mmol) and the resulting mixture was stirred at rt overnight. Upon complete consumption of starting material, the reaction mixture was added to aqueous bicarbonate solution and extracted with DCM (×2). The organic phase was passed through a phase separator and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc in heptanes to afford 160-1 (730 mg, 2.70 mmol, mixture of cis and trans isomers) as a colorless oil. The two isomers were separated via chiral SFC (Method: Column: ChiralPak IG 21×250 mm, Flow Rate: 80 g per minute cosolvent: 10% t-PrOH) to give 160-1 (300 mg, 1.11 mmol, 14% yield) Peak 1 as the trans isomer as determined with the aid of COSY, ROESY, HSQC and NOE NMR experiments. ¹H NMR (400 MHz, DMSO-d₆) δ 7.90-7.69 (m, 2H), 7.55-7.33 (m, 2H), 4.03 (d, J=7.1 Hz, 2H), 3.89-3.74 (m, 1H), 3.06 (s, 3H), 2.43 (s, 4H), 1.96-1.85 (m, 4H).

Step 2. 3-(5-(((1S,2S)-2-(ethyl(((1r,3S)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-182)

To a solution of ((1r,3r)-3-methoxycyclobutyl)methyl 4-methylbenzenesulfonate (160-1, 90 mg, 0.33 mmol) and 3-(5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-29, 82 mg, 0.22 mmol) in MeCN (1.5 mL) was added DIPEA (0.11 mL, 0.66 mmol) and the resulting mixture was stirred at 100° C. under μW irradiation for 1 h. A catalytic amount of TBAI was added and the resulting mixture was stirred at 120° C. under μW irradiation for 8 h. Additional ((1r,3r)-3-methoxycyclobutyl)methyl 4-methylbenzenesulfonate (160-1, 90 mg, 0.33 mmol) in MeCN (0.15 mL) and additional TBAI (catalytic amount) was added and stirring was continued at 120° C. under μW irradiation for 4 h. The crude reaction mixture was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 5-95% MeCN, 3.5 min gradient: Collection tubes contained several drops of formic acid) and then further purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% TFA 75 mL/min; 1.5 mL injection; Gradient: 5-95% MeCN, 3.5 min gradient) to afford TFA salt. The isolated product was dissolved in DCM and filtered through a SP—HCO₃ column eluting with DCM to afford I-182 (12 mg, 0.02 mmol, 11% yield) as a white solid. MS [M+H]⁺=470.5. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 8.06 (s, 1H), 7.70 (d, J=8.4 Hz, 1H), 7.14 (s, 1H), 7.07 (d, J=8.3 Hz, 1H), 5.13 (ddd, J=13.3, 5.2, 3.4 Hz, 1H), 4.57 (s, 1H), 4.41-4.23 (m, 2H), 3.85 (quint, J=6.4 Hz, 1H), 3.33 (s, 1H), 3.15 (s, 3H), 2.95-2.74 (m, 2H), 2.61-2.41 (m, 4H), 2.41-2.27 (m, 2H), 2.24-2.15 (m, 1H), 2.03-1.89 (m, 5H), 1.85-1.68 (m, 3H), 1.67-1.44 (m, 2H), 1.04-0.98 (m, 2H).

Example 161: Single Enantiomer of 3-(5-((2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-183)

Step 1. Single Enantiomer of 5-((2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)isobenzofuran-1(3H)-one (161-2)

8-oxa-2-azaspiro[4.5]decane hemioxalate (161-1, 200 mg, 0.537 mmol) in methanol (1 mL) was loaded onto a 1 g (0.78 mmol) SCX column. The column was flushed with methanol (×2) and eluted with 2N NH₃ in MeOH (×3). The eluant was concentrated to dryness and used in the reaction. To 154-1 (Peak 1, Example 154, 100 mg, 0.238 mmol) in MeCN (0.8 mL) was added 8-oxa-2-azaspiro[4.5]decane (161-1, 40 mg, 0.29 mmol) and DIPEA (60 μL, 0.36 mmol) and the resulting mixture was purged with nitrogen while sonicating for 20 minutes and then stirred at 120° C. for 3 h under μW radiation. The reaction mixture was added to water and extracted with dichloromethane. The combined organic phases were passed through a phase separator and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptane then 0-20% MeOH in DCM to afford 161-2 (66.3 mg, 0.185 mmol, 78% yield) as a light yellow oil. MS [M+H]⁺=358.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.81 (d, J=8.5 Hz, 1H), 7.01 (dd, J=8.5, 2.1 Hz, 1H), 6.90 (d, J=2.0 Hz, 1H), 5.25 (s, 2H), 4.88-4.55 (m, 1H), 3.63 (dt, J=8.8, 3.6 Hz, 4H), 2.95-2.35 (m, 5H), 2.24-1.96 (m, 2H), 1.86-1.42 (m, 10H).

Step 2. Single Enantiomer of Ethyl 4-((2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)-2-(chloromethyl)benzoate (161-3)

To a suspension of 161-2 (66.3 mg, 0.185 mmol) in DCE (0.9 mL) and Ethanol (0.9 mL) stirred at 70° C. was added thionyl chloride (160 μL, 2.23 mmol) dropwise and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was then cooled to rt, diluted with water, and quenched with saturated aqueous sodium bicarbonate. The mixture was extracted with EtOAc (×3) and the combined organic phases were passed through a phase separator and concentrated to afford 161-3 as a brown oil. The crude product was taken on to the next step without purification. MS [M+H]⁺=422.3

Step 3. Single Enantiomer of 3-(5-((2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-183)

To flask containing a solution of 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 60.9 mg, 0.370 mmol) in DMF (740 μL) was added DIPEA (162 μL, 0.925 mmol) and the flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at rt. for 15 minutes, then 161-3 (78.0 mg, 0.185 mmol) in DMF (1.1 mL) was added and the flask was again evacuated and backfilled with nitrogen three times. The reaction mixture was stirred at 85° C. overnight then stirred at 150° C. for 6 h under μW irradiation. The reaction mixture was then concentrated to dryness and the crude material purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane. The obtained material was then dissolved in 4:1 DCM:isopropanol and washed with saturated aqueous sodium bicarbonate. The aqueous layer was extracted twice more with 4:1 DCM:isopropanol. The combined organic phases were concentrated to dryness to afford I-183 (30.7 mg, 0.064 mmol, 35% yield) as a pale purple solid. MS [M+H]⁺=468.5. ¹H NMR (400 MHz, MeCN-d₃) δ 8.94 (s, 1H), 7.53 (dd, J=8.4, 1.6 Hz, 1H), 6.97 (d, J=2.2 Hz, 1H), 6.90 (dd, J=8.4, 2.2 Hz, 1H), 4.94 (ddd, J=13.4, 5.1, 2.1 Hz, 1H), 4.61-4.52 (m, 1H), 4.31-4.10 (m, 2H), 3.44 (q, J=5.3 Hz, 4H), 2.78-2.61 (m, 3H), 2.61-2.46 (m, 2H), 2.41 (dd, J=9.3, 1.0 Hz, 1H), 2.37-2.21 (m, 2H), 2.10-1.97 (m, 3H), 1.68-1.55 (m, 3H), 1.54-1.33 (m, 7H).

Example 162: 3-(1-oxo-5-(((1S,2S)-2-thiomorpholinocyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-184)

Step 1. Bis(2-bromoethyl)sulfane (162-2)

To a stirred solution of 2,2′-thiobis(ethan-1-ol) (162-1, 2.50 g, 20.5 mmol) in THF (50 mL) was added PBr₃ (1.35 mL, 14.3 mmol) at 15° C. and the resulting mixture was stirred at rt for 16 h. The reaction mixture was filtered and the filtrate concentrated to dryness. The obtained crude material was purified via silica gel chromatography eluting with 7% EtOAc in hexanes to afford 162-2 (1.41 g, 5.69 mmol, 28% yield) as a colorless liquid. ¹H NMR (300 MHz, CDCl₃): δ 3.48 (t, J=7.2 Hz, 4H), 2.98 (t, J=7.8 Hz, 4H).

Step 2. 3-(1-oxo-5-(((1S,2S)-2-thiomorpholinocyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-184)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 200 mg, 0.582 mmol) in MeCN (10 mL), was added bis(2-bromoethyl)sulfane (162-2, 433 mg, 1.75 mmol) and DIPEA (0.62 mL, 3.5 mmol) and the resulting mixture stirred at 120° C. for 16 h under μW irradiation. The reaction mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified via silica gel chromatography eluting with 5% MeOH in DCM to afford I-184 (95.0 mg, 0.221 mmol, 38% yield) as brown solid. MS [M+H]⁺=430.1. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.19 (s, 1H), 7.07 (dd, J=8.8, 2.0 Hz, 1H), 5.09 (dd, J=13.2, 4.8 Hz, 1H), 4.40-4.23 (m, 3H), 2.95-2.86 (m, 1H), 2.64-2.42 (m, 5H), 2.19-2.07 (m, 4H), 2.06-1.98 (m, 2H), 1.75-1.64 (m, 3H), 1.35-1.143 (m, 4H).

Example 163: 3-(5-(((1S,2S)-2-(1,4-oxazepan-4-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-185)

Step 1. 2-(benzyloxy)ethyl 4-methylbenzenesulfonate (163-2)

To a solution of 2-(benzyloxy)ethan-1-ol (163-1, 2.00 g, 13.1 mmol) in MeCN (10 mL) at 0° C. was added TEA (3.6 mL, 19 mmol), DMAP (48 mg, 0.39 mmol) and 4-methylbenzenesulfonyl chloride (3.75 g, 19.7 mmol) sequentially and the resulting mixture was stirred at rt for 3 h. The reaction mixture was filtered and the filtrate was concentrated to dryness. The obtained crude material was purified via silica gel chromatography eluting with 15% EtOAc in hexanes to afford 163-2 (3.41 g, 11.1 mmol, 85% yield) as a white solid. MS [M+H]⁺=307.3.

Step 2. 3-(2-(benzyloxy)ethoxy)propan-1-ol (163-3)

To a solution of 2-(benzyloxy)ethyl 4-methylbenzenesulfonate (163-2, 2.20 g, 7.18 mmol) in xylenes (40 mL) was added propane-1,3-diol (1.64 g, 21.5 mmol) and KOH (1.20 g, 21.5 mmol) and the resulting mixture was stirred at 150° C. for 16 h. The reaction mixture was diluted with DCM and washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified via silica gel chromatography eluting with 4% MeOH in DCM to afford 163-3 (1.20 g, 5.70 mmol, 79% yield) as a colorless oil. MS [M+H]⁺=211.3.

Step 3. 3-(2-hydroxyethoxy)propan-1-ol (163-4)

To a solution of 3-(2-(benzyloxy)ethoxy)propan-1-ol (163-3, 1.20 g, 5.71 mmol) in EtOH-EtOAc (4:1, 12 mL) under an inert atmosphere was added 10% Pd/C (240 mg). The resulting mixture was stirred under an atmosphere of hydrogen at rt for 16 h. The reaction mixture was filtered and the filtrate was concentrated to dryness to afford 163-4 (0.60 g, 4.50 mmol, 88% yield) which was carried on to the next step without purification. ¹H NMR (400 MHz, DMSO-d₆): δ 4.56 (t, J=5.2 Hz, 1H), 4.38 (t, J=5.2 Hz, 1H), 3.49-3.34 (m, 8H), 1.6-1.61 (m, 2H).

Step 4. 3-(2-(tosyloxy)ethoxy)propyl 4-methylbenzenesulfonate (163-5)

To a solution of 3-(2-hydroxyethoxy)propan-1-ol (163-4, 100 mg, 0.830 mmol) in MeCN (10 mL) at 0° C. was added TEA (0.46 mL, 3.3 mmol), DMAP (3 mg, 0.02 mmol) and 4-methylbenzenesulfonyl chloride (354 mg, 1.83 mmol) sequentially and the resulting mixture was stirred at rt for 16 h. The reaction mixture was filtered and the filtrate was concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 30% EtOAc in hexanes to afford 163-5 (230 mg, 0.540 mmol, 65% yield) as an off-white solid. MS [M+H₂O]⁺⁼445.9. ¹H NMR (300 MHz, CDCl₃): δ 7.78 (d, J=11.2 Hz, 4H), 7.33 (d, J=11.2 Hz, 4H), 4.06 (m, 4H), 3.49 (m, 4H), 2.45 (s, 6H), 1.82 (m, 2H).

Step 5. 3-(5-(((1S,2S)-2-(1,4-oxazepan-4-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-185)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 150 mg, 0.440 mmol) in MeCN (5 mL), was added 3-(2-(tosyloxy)ethoxy)propyl 4-methylbenzenesulfonate (163-5, 224 mg, 0.530 mmol) and DIPEA (0.46 mL, 2.6 mmol) and the resulting mixture was stirred at 120° C. for 16 h under μW irradiation. After complete consumption of the starting material, the reaction mixture was diluted with saturated sodium bicarbonate solution and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The obtained crude material was purified via silica gel chromatography eluting with 5% MeOH in DCM to afford I-185 (70.0 mg, 0.160 mmol, 38% yield) as a brown solid. MS [M+H]⁺=428.2. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.60 (d, J=8.8 Hz, 1H), 7.25 (s, 1H), 7.05 (dd, J=8.8, 2.0 Hz, 1H), 5.10-5.02 (m, 1H), 4.69-4.65 (m, 1H), 4.38-4.26 (m, 2H), 3.68-3.58 (m, 4H), 3.23-3.19 (m, 1H), 2.92-2.90 (m, 1H), 2.69-2.65 (m, 4H), 2.49-2.35 (m, 2H), 2.05-1.92 (m, 4H), 1.78-1.45 (m, 5H).

Example 164: 3-(5-(((1S,2S)-2-(4-(cyclopropylmethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-186)

To a solution of 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-96, 50.0 mg, 0.117 mmol) and AgOTf (164-2, 150 mg, 0.585 mmol) in DCM (0.5 mL) was added (bromomethyl)cyclopropane (164-1, 0.11 mL, 1.2 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was filtered and washed with 50% saturated aqueous Na₂CO₃. The phases were separated and the aqueous layer was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-186 (9.0 mg, 0.017 mmol, 14% yield) as an off-white solid. MS [M+H]⁺=482.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.74-7.56 (m, 1H), 7.18 (s, 1H), 7.13-6.98 (m, 1H), 5.07 (dd, J=13.2, 5.3 Hz, 1H), 4.74-4.65 (m, 1H), 4.38 (dd, J=17.2, 9.7 Hz, 1H), 4.25 (dd, J=17.1, 8.4 Hz, 1H), 3.96 (quint, J=7.2 Hz, 1H), 3.27-3.19 (m, 1H), 2.96-2.80 (m, 2H), 2.80-2.68 (m, 2H), 2.62-2.54 (m, 1H), 2.46-2.35 (m, 1H), 2.27-2.03 (m, 6H), 2.03-1.85 (m, 2H), 1.86-1.52 (m, 8H), 1.52-1.28 (m, 3H).

Example 165: 3-(5-(((1S,2S)-2-(4-isopropoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-187)

To a solution of 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-96, 50.0 mg, 0.117 mmol) and AgOTf (164-2, 150 mg, 0.585 mmol) in DCM (0.5 mL) was added 2-Bromopropane (165-1, 0.110 mL, 1.17 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was filtered and washed with 50% saturated aqueous Na₂CO₃. The phases were separated and the aqueous layer was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-187 (14.0 mg, 0.027 mmol, 23% yield) as a pale brown solid. MS [M+H]⁺=470.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.60 (d, J=8.3 Hz, 1H), 7.20-7.15 (m, 1H), 7.03 (d, J=8.6 Hz, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.73-4.65 (m, 1H), 4.39 (dd, J=17.1, 9.9 Hz, 1H), 4.25 (dd, J=17.1, 8.5 Hz, 1H), 3.70-3.61 (m, 1H), 2.96-2.81 (m, 2H), 2.80-2.67 (m, 2H), 2.64-2.54 (m, 1H), 2.44-2.35 (m, 1H), 2.21-2.02 (m, 3H), 2.02-1.84 (m, 2H), 1.80-1.69 (m, 2H), 1.69-1.55 (m, 3H), 1.54-1.40 (m, 1H), 1.41-1.28 (m, 2H), 1.20-1.13 (m, 1H), 1.04 (d, J=5.9 Hz, 6H).

Example 166: 3-(5-(((1S,2S)-2-((3aR,4R,7S,7aS)-octahydro-2H-4,7-epoxyisoindol-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-188)

Step 1. ((1R,2R,3S,4S)-7-oxabicyclo[2.2.1]heptane-2,3-diyl)dimethanol (166-2)

To a solution of Norcantharidin (166-1, 1.00 g, 5.95 mmol) in THF (20 mL) under an atmosphere of nitrogen was added LiAlH₄ (8.9 mL, 18 mmol) dropwise at 0° C. and the resulting mixture was stirred for 30 min at 0° C. and then at rt overnight. The reaction mixture was cooled to 0° C., diluted with THF (20 mL) and Na₂SO₄.10H₂O (1.50 g) was added slowly portionwise. After stirring for 10 min, Na₂SO₄ (approx. 500 mg) was added. The reaction mixture was stirred vigorously for 10 min and then the rest of Na₂SO₄.10H₂O (2.30 g) was added portionwise. Additional Na₂SO₄ (approx. 500 mg) was added and the reaction mixture was stirred for 1 hour at rt, filtered through a pad of Celite® washing with minimal amount of EtOAc. The filtrate was concentrated to dryness to afford 166-2 (623 mg, 3.94 mmol, 66% yield) as colorless oil, which was carried on to the next step without purification. ¹H NMR (400 MHz, Chloroform-d) δ 4.32 (t, J=2.7 Hz, 2H), 3.85-3.77 (m, 2H), 3.77-3.65 (m, 2H), 2.28-2.13 (m, 2H), 1.77-1.67 (m, 2H), 1.57-1.50 (m, 2H).

Step 2. ((1R,2R,3S,4S)-7-oxabicyclo[2.2.1]heptane-2,3-diyl)bis(methylene) dimethanesulfonate (166-3)

To ((1R,2R,3S,4S)-7-oxabicyclo[2.2.1]heptane-2,3-diyl)dimethanol (166-2, 83.0 mg, 0.525 mmol), DMAP (6 mg, 0.05 mmol), DIPEA (0.46 mL, 2.6 mmol) in DCM (2 mL) was added MsCl (0.10 mL, 1.3 mmol) in one portion at 0° C. and the resulting mixture was stirred at rt overnight. The reaction mixture was poured into cold saturated aqueous NaHCO₃ and extracted with DCM (×2). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness to afford 166-3 (153 mg, 0.487 mmol, 93% yield) as a brown oil, which was carried on to the next step without purification. ¹H NMR (400 MHz, Chloroform-d) δ 4.48 (dd, J=3.4, 2.2 Hz, 2H), 4.24-4.11 (m, 4H), 3.05 (s, 6H), 2.44-2.35 (m, 2H), 1.84-1.77 (m, 2H), 1.59-1.55 (m, 2H).

Step 3. 3-(5-(((1S,2S)-2-((3aR,4R,7S,7aS)-octahydro-2H-4,7-epoxyisoindol-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-188)

To 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 100 mg, 0.291 mmol) in DIPEA (0.30 mL, 1.7 mmol) and MeCN (1 mL) was added ((1R,2R,3S,4S)-7-oxabicyclo[2.2.1]heptane-2,3-diyl)bis(methylene) dimethanesulfonate (166-3, 110 mg, 0.349 mmol) and the resulting mixture was stirred for at 140° C. for 12 h under μW irradiation. The reaction mixture was concentrated to dryness and the crude product purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM to afford I-188 (9.30 mg, 0.019 mmol, 7% yield) as white powder. MS [M+H]⁺=466.2. ¹H NMR (400 MHz, DCM-d₂) δ 9.51 (br s, 1H), 7.67 (t, J=9.2 Hz, 1H), 7.05-6.76 (m, 2H), 5.07 (dd, J=13.1, 5.5 Hz, 1H), 4.77-4.53 (m, 1H), 4.47-4.23 (m, 1H), 4.23-4.01 (m, 3H), 3.27-3.00 (m, 2H), 2.93-2.70 (m, 3H), 2.33-1.91 (m, 8H), 1.77-1.51 (m, 6H), 1.40-1.25 (m, 2H).

Example 167: 3-(5-(((1S,2S)-2-(4-ethoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-189)

To a solution of 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-96, 75.0 mg, 0.175 mmol) and AgOTf (164-2, 225 mg, 0.877 mmol) in DCM (0.5 mL) was added bromoethane (167-1, 0.13 mL, 1.8 mmol) and the resulting mixture was stirred at rt for 3 h. The reaction mixture was filtered and washed with 50% saturated Na₂CO₃ (aq). The phases were separated and the aqueous phase was extracted with DCM. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) and then further purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-189 (5.0 mg, 9.5 μmol, 5% yield) as a white solid. MS [M+H]⁺=456.3. ¹H NMR (400 MHz, DCM-d₂) δ 8.04 (br s, 1H), 7.62 (d, J=8.3 Hz, 1H), 6.97-6.84 (m, 2H), 5.12-4.97 (m, 1H), 4.83 (br s, 1H), 4.34-4.11 (m, 2H), 3.37 (q, J=7.0 Hz, 2H), 3.29 (br s, 1H), 3.00 (br s, 1H), 2.94-2.63 (m, 4H), 2.43 (br s, 1H), 2.36-2.20 (m, 1H), 2.19-1.78 (m, 5H), 1.78-1.45 (m, 6H), 1.06 (t, J=7.0 Hz, 3H).

Example 168: 3-(5-(((1S,2S)-2-(7,8-dihydro-1,6-naphthyridin-6(5H)-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-190)

Step 1. Ethyl 2-(2-ethoxy-2-oxoethyl)nicotinate (168-2)

To a solution of 2-(2-ethoxy-2-oxoethyl)nicotinic acid (3.50 g, 16.7 mmol) in DMF (50 mL) was added NaHCO₃ (1.61 g, 19.2 mmol) and iodoethane (2.0 mL, 25 mmol) and the resulting mixture was stirred at 80° C. for 4 h. The reaction mixture was diluted with water and extracted with EtOAc (2×100 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified via silica gel chromatography eluting with 15% EtOAc in hexanes to afford 168-2 (3.30 g, 13.9 mmol, 83% yield) as a pale yellow oil. MS [M+H]⁺=237.9.

Step 2. 2-(3-(hydroxymethyl)pyridin-2-yl)ethan-1-ol (168-3)

To a solution of ethyl 2-(2-ethoxy-2-oxoethyl)nicotinate (168-2, 4.00 g, 16.9 mmol) in EtOH (50 mL) at 0° C. was added NaBH₄ (3.18 g, 84.3 mmol) and CaCl₂ (1.68 g, 15.2 mmol) and the resulting mixture was stirred at rt for 16 h. The reaction mixture was cooled to 0° C. quenched with 50% aqueous EtOH (10 mL) and concentrated to dryness. The obtained residue was diluted with EtOH, refluxed for 2 h, the solids were filtered off, and the filtrate was concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 5% MeOH in DCM to afford 168-3 (2.00 g, 13.1 mmol, 77% yield) as a yellow oil. MS [M+H]⁺=154.3.

Step 3. 2-(2-bromoethyl)-3-(bromomethyl)pyridine (168-4)

To a stirred solution of 2-(3-(hydroxymethyl)pyridin-2-yl)ethan-1-ol (168-3, 2.00 g, 13.1 mmol) in THF (50 mL) at 0° C. was added PBr₃ (3.7 mL, 39 mmol) and the resulting mixture was stirred at rt for 16 h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (2×75 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified via silica gel chromatography eluting with 1% MeOH in DCM to afford 168-4 (0.780 g, 0.484 mmol, 21%) as a brown solid. ¹H NMR (600 MHz, CDCl₃): 8.49 (d, J=4.8 Hz, 1H), 7.62 (d, J=7.8 Hz, 1H), 7.181-7.148 (m, 1H), 4.61-4.46 (m, 4H), 3.42-3.35 (m, 2H).

Step 4. 3-(5-(((1S,2S)-2-(7,8-dihydro-1,6-naphthyridin-6(5H)-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-190)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 200 mg, 0.580 mmol) in MeCN (10 mL) was added 2-(2-bromoethyl)-3-(bromomethyl)pyridine (168-4, 243 mg, 0.873 mmol) and DIPEA (0.62 mL, 3.5 mmol) and the resulting mixture was stirred at 120° C. for 16 h under μW irradiation. The reaction mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 5% MeOH in DCM to afford I-190 (45.0 mg, 970 μmol, 17% yield) as a pale yellow solid. MS [M+H]⁺=461.5. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 8.319-8.31 (m, 1H), 7.63 (d, J=8.4 Hz, 1H), 7.45 (d, J=7.2 Hz, 1H), 7.21 (s, 1H), 7.14-7.06 (m, 2H), 5.09-5.05 (m, 1H), 4.90-4.86 (m, 1H), 4.42 (d, J=17.2 Hz, 1H), 4.29 (d, J=17.2 Hz, 1H), 38 (d, J=14.8 Hz, 1H), 3.67 (d, J=15.6 Hz, 1H), 3.14-2.95 (m, 1H), 2.95-2.81 (m, 5H), 2.67-2.33 (m, 2H), 2.12-1.96 (m, 3H), 1.73-159 (m, 4H).

Example 169: 3-(5-(((1S,2S)-2-((((1s,4R)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-191)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 0.20 g, 0.52 mmol) and (1s,4s)-4-methoxycyclohexane-1-carbaldehyde (104-3, 0.11 g, 0.79 mmol) in TFE (10 mL) at 0° C. was added NaBH(OAc)₃ (0.17 g, 0.79 mmol) and the resulting mixture was stirred at rt for 16 h. After complete consumption of starting material, the reaction mixture was quenched with water and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified silica gel chromatography eluting with 4% MeOH in DCM to afford I-191 (52 mg, 0.11 mmol, 21% yield) as off white solid. MS [M+H]⁺=470.3. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.61 (d, J=8.4 Hz 1H), 7.18 (s, 1H), 7.04 (d, J=9.2 Hz, 1H), 5.09-5.06 (m, 1H), 4.55 (bs, 1H), 4.4 (dd, J=17.2 Hz, 1H), 4.28 (dd, J=17.2 Hz, 1H), 3.17 (s, 3H), 3.087 (bs, 1H), 2.90-2.86 (m, 1H), 2.56-2.38 (m, 2H), 2.38-2.33 (m, 3H), 2.10-2.09 (m, 1H), 1.99-1.88 (m, 3H), 1.75-1.615 (m, 5H), 1.48-1.32 (m, 5H), 1.23-176 (m, 3H).

Example 170: Single Enantiomer of 3-(5-((2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-192)

Step 1. Single Enantiomer of 5-((2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)isobenzofuran-1(3H)-one (170-2)

(R)-3-methoxypyrrolidine hydrochloride (170-1, 31 mg, 0.23 mmol) and Cs₂CO₃ (99 mg, 0.31 mmol) were suspended in MeCN (1.4 mL) and the resulting mixture stirred at rt. for 20 minutes. The reaction mixture was added to a flask containing 154-4 (Example 154, 80 mg, 0.19 mmol) and DIPEA (50 μL, 0.29 mmol) and then purged with nitrogen while sonicating for 20 minutes. The resulting mixture was stirred at 120° C. for 3 h under μW radiation. Additional Cs₂CO₃ (99 mg, 0.31 mmol) and (R)-3-methoxypyrrolidine hydrochloride (31 mg, 0.23 mmol) were added and stirring was continued at 120° C. for 3 h under μW radiation. The reaction mixture was quenched with water and extracted with dichloromethane. The organic phases were combined, passed through a phase separator, and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptane then 0-20% MeOH in DCM to afford 170-2 (12.3 mg, 0.0390 mmol, 20% yield) as a brown oil. MS [M+H]⁺=318.2.

Step 2. Single Enantiomer of Ethyl 2-(chloromethyl)-4-((2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)benzoate (170-3)

To a stirred suspension of 170-2 (12.3 mg, 0.0390 mmol)) in DCE (0.19 mL) and ethanol (0.190 mL) at 70° C. was added thionyl chloride (0.034 mL, 0.47 mmol) dropwise and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was then cooled to rt, diluted with water, quenched with saturated aqueous sodium bicarbonate and extracted with EtOAc (×3). The combined organic phases were passed through a phase separator and concentrated to afford 170-3 as a brown oil. The crude material was used directly in the next reaction. MS [M+H]⁺=382.2

Step 3. Single Enantiomer of 3-(5-((2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-192)

To a flask containing a solution of 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 13 mg, 0.078 mmol) in DMF (0.16 mL) was added DIPEA (34 μL, 0.20 mmol) and the flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at rt. for 15 minutes, 170-3 (15 mg, 0.039 mmol) in DMF (0.23 mL) was added and the flask was again evacuated and backfilled with nitrogen three times. The reaction mixture was stirred at 85° C. overnight, stirred at 150° C. for 6 h under μW irradiation and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane and then further purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient: Collection tubes contained ˜3 drops of formic acid). The fractions containing the desired product were combined and lyophilized to afford I-192 (2.0 mg, 3.9 μmol, 10% yield) as a white solid. MS [M+H]⁺ 428.2. ¹H NMR (400 MHz, MeCN-d₃) δ 8.74 (s, 1H), 7.54 (d, J=8.4 Hz, 1H), 6.98 (d, J=2.2 Hz, 1H), 6.92 (dd, J=8.5, 2.2 Hz, 1H), 4.95 (ddd, J=13.3, 5.2, 1.5 Hz, 1H), 4.67 (dt, J=6.8, 3.7 Hz, 1H), 4.30-4.14 (m, 2H), 3.78 (ddt, J=8.8, 6.1, 3.1 Hz, 1H), 3.11 (s, 3H), 2.86-2.78 (m, 2H), 2.73-2.58 (m, 5H), 2.57-2.50 (m, 1H), 2.31 (qd, J=13.1, 4.7 Hz, 1H), 2.15-1.98 (m, 2H), 1.94-1.88 (m, 1H), 1.67-1.49 (m, 5H). Absolute stereochemistry not determined.

Example 171: Single Enantiomer of 3-(5-((2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-193)

Step 1. Single Enantiomer of 5-((2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)isobenzofuran-1(3H)-one (171-2)

(S)-3-methoxypyrrolidine hydrochloride (171-1, 20 mg, 0.15 mmol) and Cs₂CO₃ (65 mg, 0.20 mmol) were suspended in MeCN (1.4 mL) and the resulting mixture stirred at rt for 20 minutes and then added to a flask containing 154-4 (Example 154, 52 mg, 0.12 mmol) and DIPEA (0.032 mL, 0.19 mmol). The flask was purged with nitrogen while sonicating for 20 minutes. The reaction mixture was stirred at 120° C. for 3 h under μW radiation and then quenched with water and extracted with dichloromethane. The organic phases were combined, passed through a phase separator and concentrated onto Celite®. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc in heptane then 0-20% MeOH in DCM to afford 171-2 (18 mg, 0.055 mmol, 20% yield) as pale yellow oil. MS [M+H]⁺=318.0. ¹H NMR (400 MHz, Chloroform-d) δ 7.82 (d, J=8.5 Hz, 1H), 7.01 (dd, J=8.5, 2.1 Hz, 1H), 6.93-6.85 (m, 1H), 5.26 (s, 2H), 4.92-4.55 (m, 1H), 4.02-3.66 (m, 1H), 3.29 (s, 3H), 3.08-2.85 (m, 3H), 2.82-2.53 (m, 2H), 2.27-2.17 (m, 1H), 2.13-2.03 (m, 2H), 1.95-1.72 (m, 5H).

Step 2. Single Enantiomer of Ethyl 2-(chloromethyl)-4-((2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)benzoate (171-3)

To a stirred suspension of 171-2 (17 mg, 0.055 mmol) in DCE (0.28 mL) and ethanol (0.28 mL) at 70° C. was added thionyl chloride (0.048 mL, 0.66 mmol) dropwise and the resulting mixture was stirred at 70° C. overnight. The reaction mixture was cooled to rt, diluted with water, and quenched with saturated aqueous sodium bicarbonate. The aqueous mixture was extracted with EtOAc (×3). The combined organic phases were passed through a phase separator and concentrated to afford 171-3 as a brown oil, which was used directly in next reaction without purification. MS [M+H]⁺=382.3

Step 3. Single Enantiomer of 3-(5-((2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-193)

To a flask containing a solution of 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 18.1 mg, 0.11 mmol) in DMF (0.22 mL) was added DIPEA (48 μL, 0.28 mmol) and flask was evacuated and backfilled with nitrogen three times. The resulting mixture was stirred at rt. for 15 minutes, 171-3 (21 mg, 0.055 mmol) in DMF (0.33 mL) was added and the flask was again evacuated and backfilled with nitrogen three times. The reaction mixture was stirred at 85° C. overnight, stirred at 150° C. for 6 h under μW irradiation and then concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 1% NEt₃) in heptane and then further purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient: Collection tubes contain ˜3 drops of formic acid). The fractions containing the desired product were combined and lyophilized to afford I-193 (5.5 mg, 11 μmol, 20% yield) as a white solid. MS [M+H]⁺=428.3. ¹H NMR (400 MHz, MeCN-d₃) δ 8.80 (s, 1H), 7.55 (d, J=8.4 Hz, 1H), 6.98 (d, J=2.2 Hz, 1H), 6.92 (dd, J=8.3, 2.1 Hz, 1H), 4.95 (ddd, J=13.6, 5.3, 2.3 Hz, 1H), 4.78-4.70 (m, 1H), 4.30-4.14 (m, 2H), 3.82 (tt, J=6.0, 2.9 Hz, 1H), 3.10 (s, 3H), 3.00-2.87 (m, 2H), 2.85-2.75 (m, 1H), 2.74-2.58 (m, 4H), 2.30 (qdd, J=13.4, 4.8, 2.8 Hz, 1H), 2.15-1.88 (m, 4H), 1.73-1.51 (m, 5H). Absolute stereochemistry not determined.

Example 172: 3-(5-(((1S,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-194)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 200 mg, 0.58 mmol) in MeCN (10 mL), (3,3-difluorocyclobutyl)methyl 4-methylbenzenesulfonate (53-2, 240 mg, 0.87 mmol) and DIPEA (0.32 mL, 1.7 mmol) were added and stirred at 120° C. for 16 h under μW irradiation. The reaction was diluted with saturated aqueous sodium bicarbonate solution and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over Na₂SO₄, filtered, and concentrated to dryness. The obtained crude material was purified via silica gel chromatography eluting with 6% MeOH in DCM to afford I-194 (50 mg, 0.11 mmol, 19% yield) as a pale brown solid. MS [M+H]⁺=448.25. ¹H NMR (400 MHz, DMSO-d₆): δ 10.98 (s, 1H), 7.62 (d, J=8.8 Hz, 1H), 7.17 (s, 1H), 7.04 (d, J=8.8 Hz, 1H), 5.07 (dd, J=13.2, 4.8 Hz, 1H), 4.58-4.54 (m, 1H), 4.41-4.22 (m, 2H), 3.13-3.11 (m, 1H), 2.95-2.87 (m, 2H), 2.67-2.51 (m, 5H), 2.42-2.12 (m, 6H), 1.99-1.85 (m, 2H), 1.72-1.55 (m, 2H), 1.48-1.39 (m, 1H).

Example 173: 3-(5-(((1S,2S)-2-(4-(difluoromethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-195)

To a solution of 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-96, 100 mg, 0.234 mmol) in MeCN (4 mL), whilst bubbling with nitrogen, was added 2,2-difluoro-2-(fluorosulfonyl)acetic acid (173-1, 208 mg, 1.17 mmol) and CuI (9 mg, 0.5 μmol) and the resulting mixture was stirred at 100° C. for 1 hour in the μW. The reaction mixture was added to water (20 mL) and saturated aqueous sodium hydrogen carbonate (20 mL). The aqueous mixture was extracted with 20% i-PrOH in DCM (×3) and the combined organic phases were passed through a phase separating column and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0-15% i-PrOH (with 0.1% NEt₃) in DCM (with 0.1% NEt₃) and then further purified via reverse phase HPLC (Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 10-30% MeCN, 3.5 min gradient) to afford the formate salt of I-195 (7.0 mg, 0.013 mmol, 5% yield) as a white solid. MS [M+H]⁺=478.3. ¹H NMR (400 MHz, DMSO-d₆) δ 10.95 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.18 (s, 1H), 7.03 (d, J=8.6 Hz, 1H), 6.69 (t, J=77.9 Hz, 1H), 5.06 (dd, J=13.2, 5.1 Hz, 1H), 4.74-4.67 (m, 1H), 4.45-4.19 (m, 2H), 4.15-4.02 (m, 1H), 2.98-2.82 (m, 2H), 2.80-2.65 (m, 2H), 2.59 (d, J=16.9 Hz, 1H), 2.40 (td, J=13.1, 4.5 Hz, 1H), 2.30-2.19 (m, 2H), 2.14-1.77 (m, 5H), 1.73-1.40 (m, 6H).

Example 174: 3-(5-(((1S,2S)-2-(1,3-dihydro-2H-pyrrolo[3,4-c]pyridin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-196)

Step 1. Pyridine-3,4-diyldimethanol hydrochloride (174-2)

To a solution of dimethyl pyridine-3,4-dicarboxylate (174-1, 1.50 g, 7.68 mmol) in EtOH (45 mL) at 0° C. was added NaBH₄ (1.45 g, 38.4 mmol) and CaCl₂ (0.760 g, 6.92 mmol) and the resulting mixture was stirred at rt for 16 h. The reaction mixture was cooled to 0° C., quenched with 50% aqueous EtOH (10 mL), and concentrated to dryness. The obtained residue was diluted with EtOH, refluxed for 2 h, the solids were filtered off, and the filtrate was concentrated under reduced pressure. The obtained crude material was treated with 1,4-dioxane-HCl, stirred for 1 h and concentrated to dryness. The residue was triturated with diethyl ether and dried under high vacuum to afford the HCl salt of 174-2 (1.30 g, 7.40 mmol, 96% yield) as white solid. MS [M+H]⁺=140.2.

Step 2. 3,4-Bis(chloromethyl)pyridine hydrochloride (174-3)

To a stirred solution of pyridine-3,4-diyldimethanol hydrochloride (174-2, 800 mg, 4.55 mmol) in DCM (15 mL) at 0° C. was added SOCl₂ (15 mL) and the resulting mixture was stirred at rt for 16 h. The reaction mixture was concentrated to dryness. The obtained crude material was treated with 1,4-dioxane-HCl, stirred for 1 h and concentrated to dryness. The crude material was triturated with diethyl ether and dried under high vacuum to afford the HCl salt of 174-3 (800 mg, 3.76 mmol, 83% yield) as a white solid. MS [M+H]⁺=176.2.

Step 3. 3-(5-(((1S,2S)-2-(1,3-dihydro-2H-pyrrol[3,4-c]pyridin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-196)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 150 mg, 0.440 mmol) in MeCN (10 mL) was added 3,4-bis(chloromethyl)pyridine hydrochloride (174-3, 139 mg, 0.650 mmol) and DIPEA (0.39 mL, 2.2 mmol) and the resulting mixture was stirred at 70° C. for 16 h under μW irradiation. The reaction mixture was cooled to rt and filtered. The filtrate was treated with 1,4-dioxane-HCl and stirred for 1 h. The resulting solution was lyophilized, the obtained crude material was purified by reverse phase HPLC (Column: KINETEX (150 mm×21.2 mm), Mobile phase A: 0.01% HCOOH (aq), Mobile phase B: MeCN, Time (min)/% B: 0/5, 2/10, 10/628.4 Flow rate: 20 ml/min, Diluent: Mobile phase). The fractions containing the desired product were combined and lyophilized to afford I-196 (15.0 mg, 33.0 μmol, 7% yield). MS [M+H]⁺=447.4. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 8.46 (s, 1H), 8.39-8.38 (m, 1H) 7.62 (d, J=8.4 Hz, 1H), 7.36-7.30 (m, 1H), 7.21 (s, 1H), 7.09 (d, J=8.4 Hz, 1H), 5.09-5.04 (m, 1H), 4.90-4.85 (m, 1H), 4.43-4.22 (m, 3H), 3.99-3.96 (m, 3H), 3.29-3.24 (m, 1H), 2.95-2.86 (m, 2H), 2.66-2.32 (m, 1H), 2.30-2.22 (m, 2H), 2.0-1.96 (m, 2H), 1.74-1.65 (m, 3H).

Example 175: 3-(5-(((1S,2S)-1-amino-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-197)

Step 1. tert-butyl ((1S,2S)-2-hydroxy-2,3-dihydro-1H-inden-1-yl)carbamate (175-2)

A mixture of (3aR,6S,6aS)-octahydrocyclopenta[b]pyrrol-6-ol (175-1, 2.00 g, 13.4 mmol), di-tert-butyl dicarbonate (3.51 g, 16.1 mmol), TEA (2.8 mL, 20 mmol) and MeOH (10 mL) was stirred vigorously at rt for 48 hours. The reaction mixture was diluted with brine (25 mL) and extracted with DCM (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to afford 175-2 (3.28 g, 13.2 mmol, 98% yield) as a yellow oil. The product was used in the next step without purification. ¹H NMR (400 MHz, DCM-d₂) δ 7.37-7.09 (m, 4H), 5.09 (s, 1H), 4.84 (t, J=6.2 Hz, 1H), 4.34 (td, J=7.8, 6.3 Hz, 1H), 3.97 (s, 1H), 3.23 (dd, J=15.7, 7.6 Hz, 1H), 2.82 (dd, J=15.7, 8.1 Hz, 1H), 1.46 (s, 9H).

Step 2. tert-butyl ((1S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)-2,3-dihydro-1H-inden-1-yl)carbamate (175-3)

To a mixture of 3-(5-bromo-1-oxoisoindolin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (31-3a, 2.0 g, 4.4 mmol), Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (0.049 g, 0.044 mmol), NiCl₂(glyme) (0.048 g, 0.22 mmol), TMP (1.1 mL, 6.6 mmol) and tert-butyl ((1S,2S)-2-hydroxy-2,3-dihydro-1H-inden-1-yl)carbamate (175-2, 1.21 g, 4.9 mmol) under an atmosphere of nitrogen was MeCN (20 mL) and the resulting mixture was stirred under irradiation of Blue LED lights for 20 h. The reaction mixture was filtered through a pad of Celite and concentrated to dryness. The crude material was purified via silica gel chromatography eluting with 0 to 40% EtOAc:EtOH (v/v=3:1) in DCM to afford 175-3 (2.51 g, 4.04 mmol, 92% yield) as a pale yellow solid. MS [M−H]⁻=620.2.

Step 3. 3-(5-(((1S,2S)-1-amino-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-197)

To tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)-2,3-dihydro-1H-inden-1-yl)carbamate (175-3, 1.92 g, 3.09 mmol) in DCM (7 mL) was added methanesulfonic acid (0.80 mL, 12 mmol) and the resulting mixture was stirred at rt for 20 h. TEA (4.3 mL, 31 mmol) was added dropwise at 0° C. and the reaction mixture was allowed to reach rt. N1,N2-dimethylethane-1,2-diamine (0.37 mL, 3.4 mmol) was added and stirring was continued at rt overnight. The reaction mixture was diluted with 50% saturated aqueous NaHCO₃ in water and extracted with DCM:i-PrOH (v/v=4:1) (×2). The combined organic phases were concentrated to dryness to afford crude I-197 (1.18 g, 3.09 mmol). A small amount of crude material was purified by reverse phase HPLC (Method Column: X-bridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-197 (5.0 mg, 0.01 mmol, 0.4% yield) as a white solid. MS [M+H]⁺=392.1. ¹H NMR (400 MHz, DMSO-d₆) δ 10.97 (s, 1H), 8.30 (s, 1H), 7.63 (d, J=8.4 Hz, 1H), 7.47-7.31 (m, 1H), 7.31-7.17 (m, 4H), 7.12 (dd, J=8.4, 2.2 Hz, 1H), 5.08 (dd, J=13.3, 5.1 Hz, 1H), 4.84 (q, J=4.8 Hz, 1H), 4.52-4.21 (m, 3H), 3.61-3.54 (m, 2H), 3.10-2.75 (m, 2H), 2.66-2.55 (m, 1H), 2.46-2.26 (m, 1H), 2.06-1.93 (m, 1H).

Example 176: 3-(5-(((1S,2S)-1-(ethylamino)-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-198)

To 3-(5-(((1S,2S)-1-amino-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH salt (I-197, 60 mg, 0.14 mmol) in DIPEA (0.2 mL, 0.6 mmol) and DMF (1 mL) was added iodoethane (0.016 mL, 0.20 mmol) in one portion and the resulting mixture was stirred at rt overnight. The reaction mixture was concentrated to dryness and the crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collected fractions contain a drop of formic acid) to afford the formate salt of I-198 (19 mg, 0.041 mmol, 29% yield) as a white powder. MS [M+H]⁺=420.5. ¹H NMR (400 MHz, Deuterium Oxide) δ 8.45 (s, 1H), 7.77 (d, J=8.5 Hz, 1H), 7.49 (d, J=7.1 Hz, 1H), 7.43-7.32 (m, 3H), 7.22 (s, 1H), 7.15 (d, J=8.6 Hz, 1H), 5.24-5.18 (m, 1H), 5.13 (dd, J=13.3, 4.9 Hz, 1H), 4.64-4.42 (m, 3H), 3.65 (dd, J=17.3, 5.8 Hz, 1H), 3.07 (d, J=17.2 Hz, 1H), 2.98-2.77 (m, 4H), 2.60-2.42 (m, 1H), 2.36-2.20 (m, 1H), 1.15 (t, J=7.1 Hz, 3H).

Example 177: 3-(5-(((1S,2S)-1-(diethylamino)-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-199)

To 3-(5-(((1S,2S)-1-amino-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH salt (I-197, 120 mg, 0.267 mmol), acetaldehyde (2-1, 0.06 mL, 1 mmol) in TFE (2 mL) was added NaBH(OAc)₃ (226 mg, 1.07 mmol) in one portion and the resulting mixture was stirred at rt overnight. The reaction mixture was concentrated to dryness and the crude material purified via silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-199 (18.9 mg, 0.0410 mmol, 15% yield) as a white powder. MS [M+H]⁺=448.1. ¹H NMR (400 MHz, DCM-d₂) δ 8.47 (s, 1H), 7.96-7.57 (m, 1H), 7.40-7.27 (m, 1H), 7.26-7.14 (m, 3H), 7.13-7.00 (m, 2H), 5.21-4.97 (m, 2H), 4.72 (d, J=4.0 Hz, 1H), 4.51-4.21 (m, 2H), 3.50 (dd, J=16.9, 7.3 Hz, 1H), 2.94 (dd, J=16.9, 4.4 Hz, 1H), 2.87-2.78 (m, 2H), 2.66-2.44 (m, 4H), 2.42-2.24 (m, 1H), 2.22-2.13 (m, 1H), 1.07 (t, J=7.1 Hz, 6H).

Example 178: 3-(5-(((1S,2S)-1-(ethyl(methyl)amino)-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-200) and 3-(5-(((1S,2S)-1-(dimethylamino)-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-201)

Step 1. Mixture of 3-(5-(((1S,2S)-1-amino-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1S,2S)-1-amino-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)-1-(hydroxymethyl)piperidine-2,6-dione (178-1)

To tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)-2,3-dihydro-1H-inden-1-yl)carbamate (175-3, 1.92 g, 3.09 mmol) in DCM (7 mL) was added methanesulfonic acid (0.80 mL, 12 mmol) and the resulting mixture was stirred at rt for 20 hr. TEA (4.3 mL, 31 mmol) was added dropwise at 0° C. and the reaction mixture was allowed to reach rt. N1,N2-dimethylethane-1,2-diamine (0.37 mL, 3.4 mmol) was added and stirring was continued at rt overnight. The reaction mixture was diluted with 50% saturated aqueous NaHCO₃ in water and extracted with DCM:i-PrOH (v/v=4:1) (×2). The combined organic phases were concentrated to dryness to afford 178-1 (1.18 g, 3.09 mmol), which was carried on to the next step without purification.

Step 2. 3-(5-(((1S,2S)-1-(ethyl(methyl)amino)-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-200) and 3-(5-(((1S,2S)-1-(dimethylamino)-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-201)

To a mixture of 3-(5-(((1S,2S)-1-amino-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(5-(((1S,2S)-1-amino-2,3-dihydro-1H-inden-2-yl)oxy)-1-oxoisoindolin-2-yl)-1-(hydroxymethyl)piperidine-2,6-dione (178-1, 100 mg, 0.222 mmol) and NaBH(OAc)₃ (141 mg, 0.667 mmol) in DCM (2 mL) under an atmosphere of nitrogen was added acetaldehyde (2-1, 0.037 mL, 0.67 mmol) in one portion and the resulting mixture was stirred for at rt for 2 days and then concentrated to dryness. The crude material was purified by reverse phase HPLC (Column: Xbridge C18 OBD 5 um 30×50 mm; Conditions: Water/MeCN with 5 mM NH₄OH 75 mL/min; 1.5 mL injection; Gradient: 15-40% MeCN, 3.5 min gradient; collected fractions contain a drop of formic acid) to afford the formic acid salt I-201 (13.0 mg, 0.028 mmol, 12% yield) was obtained as a white powder.

The fractions that contained impure I-200 were combined, lyophilized and then further via silica gel chromatography eluting with 0-100% EtOAc:EtOH (v/v=3:1, with 0.1% NEt₃) in DCM (with 0.1% NEt₃) to afford I-200 (8.9 mg, 0.021 mmol, 9% yield) as a white powder. Data for I-200: MS [M+H]⁺=434.2. ¹H NMR (400 MHz, DCM-d₂) δ 8.21 (s, 1H), 7.73 (d, J=8.2 Hz, 1H), 7.37-7.30 (m, 1H), 7.28-7.17 (m, 3H), 7.11-7.04 (m, 2H), 5.19-5.05 (m, 2H), 4.57 (d, J=3.5 Hz, 1H), 4.42-4.26 (m, 2H), 3.49 (dd, J=17.0, 7.0 Hz, 1H), 2.96 (dd, J=17.0, 3.9 Hz, 1H), 2.90-2.73 (m, 2H), 2.56 (q, J=7.1 Hz, 2H), 2.41-2.27 (m, 1H), 2.20 (s, 3H), 2.19-2.13 (m, 1H), 1.10 (t, J=7.1 Hz, 3H). Data for I-201: MS [M+H]⁺=420.2. ¹H NMR (400 MHz, DCM-d₂) δ 8.20 (s, 1H), 8.11 (s, 1H), 7.74 (d, J=8.4 Hz, 1H), 7.47 (d, J=7.2 Hz, 1H), 7.37-7.22 (m, 3H), 7.14-7.09 (m, 1H), 7.05 (dd, J=8.4, 2.2 Hz, 1H), 5.27-5.19 (m, 1H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.61 (d, J=3.0 Hz, 1H), 4.43-4.28 (m, 2H), 3.54 (dd, J=17.1, 6.7 Hz, 1H), 3.02 (dd, J=17.1, 3.3 Hz, 1H), 2.90-2.74 (m, 2H), 2.40 (s, 6H), 2.38-2.25 (m, 1H), 2.24-2.14 (m, 1H).

Example 179: 3-(5-(((1R,2S,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-207) and 3-(5-(((1R,2R,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-208)

Step 1. Diethyl 3-methylhexanedioate (179-2)

To a solution of 3-methyladipic acid (179-1, 2.00 g, 12.6 mmol) in ethanol (20 mL) was added thionyl chloride (2.0 mL, 27 mmol) dropwise and the resulting mixture was stirred for 1 hour at room temperature. The reaction solution was diluted with a 1:1 solution of sat. sodium bicarbonate solution (50 mL) and extracted with dichloromethane (3×50 mL). The combined organic phases were then dried over magnesium sulfate, filtered, and concentrated to afford crude 186-2 (2.71 g, 12.5 mmol). The crude product was carried onto the next step without purification. MS [M+H]⁺=217.2. ¹H NMR (400 MHz, Chloroform-d) δ 4.22-4.09 (m, 4H), 2.45-2.26 (m, 3H), 2.23-2.09 (m, 1H), 1.81-1.65 (m, 1H), 1.62-1.47 (m, 1H), 1.36 (t, J=7.1 Hz, 1H), 1.33-1.19 (m, 6H), 0.98 (d, J=6.7 Hz, 3H).

Step 2. Ethyl 2-methyl-5-oxocyclopentane-1-carboxylate (179-3)

To a solution of diethyl 3-methylhexanedioate (179-2, 2.71 g, 12.5 mmol) in THF (60 mL) was added sodium ethoxide (5.70 mL, 15.3 mmol) and the resulting mixture was stirred at 70° C. overnight. The reaction solution was quenched with water (40 mL) and then extracted with dichloromethane (3×40 mL), The combined organic phases were dried over magnesium sulfate, filtered, and concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-40% EtOAc in heptanes to afford 179-3 (1.42 g, 8.31 mmol, 66% yield mixture of regioisomers) as a liquid. MS [M+H]⁺=171.1.

Step 3. Ethyl trans-2-hydroxy-5-methylcyclopentane-1-carboxylate (179-4)

To a solution of ethyl 2-methyl-5-oxocyclopentane-1-carboxylate (179-3, 1.42 g, 8.31 mmol) in Et₂O (28 mL) was added zinc(II) tetrahydroborate (150-2, 24 mL, 9.60 mmol, 0.4M in THF) and the resulting mixture was stirred at 0° C. for 1 hour. The reaction mixture was then quenched dropwise with sat. ammonium acetate solution (100 mL) and extracted with dichloromethane (3×80 mL). The combined organic phases were then dried over magnesium sulfate, filtered, and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0-20% EtOAc in heptane to afford 179-4 (496 mg, 2.88 mmol, 35% yield, mixture of regioisomers and diastereomers) as a colorless liquid. MS [M+H]⁺=173.2.

Step 4. Ethyl trans-2-((tert-butyldiphenylsilyl)oxy)-5-methylcyclopentane-1-carboxylate (179-5)

To a solution of ethyl trans-2-hydroxy-5-methylcyclopentane-1-carboxylate (179-4, 496.4 mg, 2.88 mmol), imidazole (498 mg, 7.32 mmol), and DMAP (31.7 mg, 0.259 mmol) in THF (10 mL) was added TBDPSCl (0.90 mL, 3.5 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was diluted with ethyl acetate (160 mL) and washed with water (30 mL), saturated aqueous sodium bicarbonate solution (30 mL), and brine (20 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0-10% EtOAc in heptane to afford 179-5, as a clear, viscous liquid. This sample was then further purified using chiral SFC (Column: 2.1×25.0 cm Chiralpak OD-H; CO₂ co-solvent: heptane/2-propanol (v/v=9:1) with 10 mM NH₃; Isocratic method: 15% co-solvent at 80 g/min; System pressure: 150 bar) to afford 179-5 (513 mg, 1.19 mmol, 41% yield, mixture of trans-regioisomers) as a colorless oil. Chiral SFC Peak 1: Rt 0.83 mins; MS [M+18]⁺=428.4.

Step 5. trans-2-((tert-butyldiphenylsilyl)oxy)-5-methylcyclopentane-1-carboxylic Acid (179-6)

Ethyl trans-2-((tert-butyldiphenylsilyl)oxy)-5-methylcyclopentane-1-carboxylate (179-5, 513 mg, 1.25 mmol) and lithium hydroxide monohydrate (519.9 mg, 12.39 mmol) were dissolved in THF (3 mL), MeOH (3 mL), and Water (1 mL) in a reaction vial with stir bar and the resulting solution was stirred at rt for 5 days. The reaction mixture was diluted with water (30 mL) and acidified with 1 N HCl solution to ˜pH 5. The solution was then extracted with ethyl acetate (3×50 mL). The combined organic phases were then dried over magnesium sulfate, filtered, and concentrated to dryness to afford crude 179-6 (591 mg, 1.54 mmol, mixture of trans-regioisomers) as a yellow oil that was used in the next step without purification. MS [M]⁻=381.3.

Step 6. tert-butyl (trans-2-((tert-butyldiphenylsilyl)oxy)-5-methylcyclopentyl)carbamate (179-7)

To a reaction vial containing trans-2-((tert-butyldiphenylsilyl)oxy)-5-methylcyclopentane-1-carboxylic acid (179-6, 478 mg, 1.25 mmol), TBAB (60 mg, 0.19 mmol), and zinc trifluoromethanesulfonate (23 mg, 0.062 mmol, 0.4M in THF) was added sodium azide (298 mg, 4.58 mmol) and THF (4 mL) and the reaction vial was placed under an atmosphere of nitrogen. Boc-anhydride (320 mg, 1.47 mmol) and tert-butanol (0.072 mL, 0.75 mmol) were then added and the resulting mixture was stirred at 40° C. for 24 hours. The reaction mixture was quenched with 10% sodium nitrite solution (25 mL) and ethyl acetate (10 mL) and stirring was continued at rt for 30 minutes. The solution was diluted with ethyl acetate (120 mL) and washed with saturated aq. sodium bicarbonate solution (2×20 mL) and brine (20 mL). The organic phase was dried over magnesium sulfate, filtered, and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0-20% EtOAc in heptane to afford 179-7 (274 mg, 0.603 mmol, 48% yield) as a clear liquid, as a mixture of trans-regioisomers. MS [M+H]⁺=454.4.

Step 7. tert-butyl (trans-2-hydroxy-5-methylcyclopentyl)carbamate (179-8)

To a reaction vial with stir bar containing a solution of tert-butyl (trans-2-((tert-butyldiphenylsilyl)oxy)-5-methylcyclopentyl)carbamate (179-7, 162 mg, 0.358 mmol) in THF (2 mL) was added 1M TBAF in THF (0.45 mL, 0.45 mmol) and the resulting mixture was stirred at rt overnight. The reaction solution was concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-50% EtOAc in heptane to afford 179-8 (92.1 mg, 0.342 mmol, 96% yield) as a white solid, as a mixture of trans-regioisomers. MS [M+H]⁺=216.3.

Step 8. tert-butyl ((1R,2S,5R)-2-methyl-5-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate (179-10) and tert-butyl ((1S,2R,5S)-2-methyl-5-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate (179-11)

To a reaction vial with stir bar was added 5-bromoisobenzofuran-1(3H)-one (179-9, 89.8 mg, 0.422 mmol), tert-butyl (trans-2-hydroxy-5-methylcyclopentyl)carbamate (179-8, 92.1 mg, 0.428 mmol), NiCl₂(glyme) (4.5 mg, 0.020 mmol), dtbbpy (10 mg, 0.038 mmol), and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (4.6 mg, 4.1 μmol) and the reaction vial was then purged with nitrogen gas. MeCN (2 mL) and 2,2,6,6-tetramethylpiperidine (0.08 mL, 0.5 mmol) were added and the resulting mixture was stirred under blue LED lights at rt over 3 d. The crude solution was diluted with ethyl acetate and filtered through Celite®. The precipitate was washed with ethyl acetate and the filtrate concentrated to dryness. The crude material was purified by silica gel chromatography eluting with 0-55% EtOAc in heptane to provide the racemic trans products. The mixture was separated by chiral SFC (Column: 2.1×25.0 cm Chiralpak IH; CO₂ co-solvent: IPA; Isocratic method: 35% co-solvent at 80 g/min; System pressure: 150 bar) to afford: Enantiomer 1 (179-10) (Chiral SFC Rt=2.07 mins, 24.0 mg, 0.068 mmol, 16% yield) as a white solid, MS [M-tBu+H]⁺=292.3 ¹H NMR (400 MHz, Methylene Chloride-d) δ 7.75 (d, J=8.5 Hz, 1H), 7.04 (dd, J=8.5, 2.1 Hz, 1H), 7.00-6.90 (m, 1H), 5.21 (s, 2H), 4.71-4.50 (m, 2H), 3.61 (td, J=8.5, 5.0 Hz, 1H), 2.19-2.04 (m, 1H), 1.97-1.75 (m, 3H), 1.53-1.46 (m, 1H), 1.41 (s, 9H), 1.11 (d, J=6.5 Hz, 3H): Enantiomer 2 (179-11) (Chiral SFC Rt=2.30 mins, 25.0 mg, 0.071 mmol, 17% yield) as a white solid, MS [M-tBu+H]⁺=292.3. ¹H NMR (400 MHz, Methylene Chloride-d) δ 7.75 (d, J=8.5 Hz, 1H), 7.04 (dd, J=8.5, 2.1 Hz, 1H), 6.96 (s, 1H), 5.21 (s, 2H), 4.74-4.49 (m, 2H), 3.61 (td, J=8.5, 4.9 Hz, 1H), 2.21-2.03 (m, 1H), 1.98-1.73 (m, 3H), 1.52-1.47 (m, 1H), 1.41 (s, 9H), 1.11 (d, J=6.5 Hz, 3H). The absolute stereochemistry of the two enantiomers corresponding to the two product peaks is unknown and was assigned arbitrarily.

Step 9. 5-(((1R,2R,3S)-2-amino-3-methylcyclopentyl)oxy)isobenzofuran-1(3H)-one (179-12)

To a reaction vial with stir bar containing a solution of tert-butyl ((1R,2S,5R)-2-methyl-5-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate (179-10, Peak 1, 24 mg, 0.069 mmol) in DCM (1 mL) was added TFA (0.05 mL, 0.65 mmol) and the resulting solution was stirred at rt for 14 h. The solution was then concentrated to dryness. The obtained product was then azeotroped with methanol and dichloromethane to afford the crude 179-12 as a clear gum which was carried on to the next step without purification MS [M+H]⁺=248.3.

Step 10. 5-(((1R,2R,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)isobenzofuran-1(3H)-one (179-13)

To a reaction vial with stir bar containing a solution of 5-(((1R,2R,3S)-2-amino-3-methylcyclopentyl)oxy)isobenzofuran-1(3H)-one (179-12, 25 mg, 0.069 mmol) and acetaldehyde (0.02 mL, 0.356 mmol) in DMF (0.5 mL) was added sodium triacetoxyborohydride (78 mg, 0.368 mmol) in one portion and the resulting mixture was stirred at rt for 23 h. The reaction mixture was diluted dichloromethane and 50% sat. aqueous sodium bicarbonate solution (10 mL) and passed through a phase separator. The aqueous solution was extracted with dichloromethane. The combined organic phases were then concentrated to dryness to afford crude 179-13 as an amber liquid. MS [M+H]⁺=304.3. The product was carried through to the next step without purification.

Step 11. Ethyl 2-(chloromethyl)-4-(((1R,2R,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)benzoate (179-14)

To a solution of 5-(((1R,2R,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)isobenzofuran-1 (3H)-one (179-13, 21 mg, 0.069 mmol) in ethanol (1 mL) was added thionyl chloride (0.03 mL, 0.4 mmol) and the resulting mixture was stirred at 70° C. for 17 h. The reaction mixture was concentrated and azeotroped with dichloromethane to afford crude 179-14 as a light brown solid. The product was carried through to the next step without purification. MS [M+H]⁺=368.3.

Step 12. 3-(5-(((1R,2R,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione HC(O)OH Salt (I-207)

To a solution of ethyl 2-(chloromethyl)-4-(((1R,2R,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)benzoate (179-14, 25.5 mg, 0.069 mmol) in DMF (0.5 mL) and DIPEA (0.07 mL, 0.4 mmol) sparged with nitrogen was added 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 18.8 mg, 0.114 mmol) in one portion and the resulting mixture was sparged with nitrogen again. The reaction mixture was stirred at 110° C. for 20 h and then diluted with ethyl acetate (40 mL). The aqueous mixture was washed with sat. aqueous sodium bicarbonate solution (2×10 mL) and brine (10 mL), dried over magnesium sulfate, filtered and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0-100% EtOH:EtOAc (v/v=1:3) in DCM with 0.1% triethylamine modifier and then further purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 m 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient) to afford the formate salt of I-207 (0.5 mg, 1 μmol, 2% yield). MS [M+H]⁺=414.4. ¹H NMR (400 MHz, Acetonitrile-d₃) δ 8.79 (s, 1H), 8.10 (s, 1H), 7.63 (d, J=8.4 Hz, 1H), 7.13-7.06 (m, 1H), 7.01 (dd, J=8.4, 2.2 Hz, 1H), 5.11-4.94 (m, 1H), 4.85-4.73 (m, 1H), 4.43-4.22 (m, 2H), 3.02 (dd, J=9.3, 5.0 Hz, 1H), 2.89-2.58 (m, 7H), 1.75-1.66 (m, 1H), 1.37 (dtd, J=12.4, 10.2, 7.7 Hz, 1H), 1.12 (d, J=6.5 Hz, 3H), 1.06 (t, J=7.1 Hz, 6H).

Step 13. 5-(((1S,2R,5S)-2-amino-3-methylcyclopentyl)oxy)isobenzofuran-1(3H)-one (179-15)

To a reaction vial with stir bar containing a solution of tert-butyl ((1S,2R,5S)-2-methyl-5-((1-oxo-1,3-dihydroisobenzofuran-5-yl)oxy)cyclopentyl)carbamate (179-11, Peak 2, 25 mg, 0.072 mmol) in DCM (1 mL) was added TFA (0.05 mL, 0.65 mmol) and the resulting solution was stirred at rt for 14 h. The reaction mixture was concentrated to dryness and then azeotroped with methanol and dichloromethane to afford the crude 179-15 as a clear gum. The product was carried through to the next step without purification. MS [M+H]⁺=248.3.

Step 14. 5-(((1S,2s,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)isobenzofuran-1(3H)-one (179-16)

To a reaction vial with stir bar containing a solution of 5-(((1S,2R,5S)-2-amino-3-methylcyclopentyl)oxy)isobenzofuran-1(3H)-one (179-15, 26 mg, 0.072 mmol) and acetaldehyde (0.02 mL, 0.4 mmol) in DMF (0.5 mL) was added sodium triacetoxyborohydride (82 mg, 0.39 mmol) in one portion and the resulting mixture was stirred at room temperature for 23 h. The reaction solution was diluted dichloromethane and 50% sat. aqueous sodium bicarbonate solution (10 mL) and passed through a phase separator. The aqueous solution was extracted with dichloromethane and then concentrated to dryness to afford crude 179-16 (25 mg, 0.065 mmol, 90% yield) as an amber liquid. The product was carried through to the next step without purification. MS [M+H]⁺=304.4.

Step 15. Ethyl 2-(chloromethyl)-4-(((1S,2S,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)benzoate (179-17)

To a solution of 5-(((1S,2S,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)isobenzofuran-1(3H)-one (179-16, 22 mg, 0.073 mmol) in EtOH (1 mL) was added thionyl chloride (0.03 mL, 0.4 mmol) and the resulting mixture was stirred at 70° C. for 17 h. The reaction solution was concentrated and azeotroped with dichloromethane to afford crude 179-17 as a dark brown gum. The crude material was taken through to the next reaction step without purification. MS [M+H]⁺=368.3.

Step 16. 3-(5-(((1S,2S,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-208)

To a solution of ethyl 2-(chloromethyl)-4-(((1S,2S,3S)-2-(diethylamino)-3-methylcyclopentyl)oxy)benzoate (179-17, 26.7 mg, 0.073 mmol) in DMF (0.5 mL) and DIPEA (0.07 mL, 0.4 mmol) and sparged with nitrogen was added 3-aminopiperidine-2,6-dione hydrochloride (I-1c, 19.5 mg, 0.118 mmol) in one portion and the resulting mixture was again sparged with nitrogen. The reaction mixture was stirred at 110° C. for 20 h and then diluted with ethyl acetate (40 mL). The mixture was washed with sat. aqueous sodium bicarbonate solution (2×10 mL) and brine (10 mL), dried over magnesium sulfate, filtered, and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0-100% EtOH:EtOAc (v/v=1:3) in DCM with 0.1% triethylamine modifier and then further purified by reverse phase HPLC (Column: X-bridge C18 OBD 5 m 30×50 mm; Conditions: Water/MeCN with 0.1% Formic Acid 75 mL/min; 1.5 mL injection; Gradient: 5-20% MeCN, 3.5 min gradient). The fractions containing the desired product were combined and lyophilized to afford I-208 (1.1 mg, 2.27 μmol, 3% yield): MS [M+H]⁺=414.4. ¹H NMR (400 MHz, DMSO-d₆) δ 10.96 (s, 1H), 7.60 (d, J=8.5 Hz, 1H), 7.20-7.14 (m, 1H), 7.07-6.98 (m, 1H), 5.12-5.01 (m, 1H), 4.76-4.66 (m, 1H), 4.44-4.22 (m, 2H), 2.96-2.82 (m, 2H), 2.06-1.92 (m, 2H), 1.89-1.71 (m, 3H), 1.66-1.53 (m, 1H), 1.38-1.25 (m, 2H), 1.05 (d, J=6.3 Hz, 3H), 0.99 (t, J=7.0 Hz, 6H).

Example 180: 3-(5-(((1S,2S)-2-(3-ethoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-202)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 200 mg, 0.580 mmol) in acetonitrile (5 mL) was added 2-ethoxypropane-1,3-diyl bis(4-methylbenzenesulfonate) (54-1, 374 mg, 0.870 mmol) and DIPEA (0.36 mL, 1.7 mmol) and the resulting mixture was stirred at 120° C. for 16 h under microwave irradiation. The reaction mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted with DCM (3×30 mL). The combined organic extracts were washed with brine, dried over Na₂SO₄ and concentrated to dryness. The obtained crude material was purified by silica gel chromatography eluting with 6% MeOH in DCM to afford I-202 (25.0 mg, 0.0580 mmol, 10% yield) as a white solid. MS [M+H]⁺=428.25. ¹H NMR (400 MHz, DMSO-d₆): δ 10.97 (s, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.13 (s, 1H), 7.02 (d, J=8.4 Hz, 1H), 5.09 (dd, J=13.2, 4.8 Hz, 1H), 4.52 (bs, 1H), 4.40-4.23 (m, 2H), 4.00-3.97 (m, 1H), 3.52-3.56 (m, 2H), 2.87-2.810 (m, 5H), 2.60-2.32 (m, 2H), 2.10-1.96 (m, 3H), 1.75-1.63 (m, 4H), 1.39-1.38 (m, 1H), 1.07 (t, 6.8 Hz, 3H).

Example 181: 3-(1-oxo-5-(((1S,2S)-2-(3-(pyridin-3-yloxy)azetidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione (I-203)

To a solution of 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-12, 200 mg, 0.580 mmol) in acetonitrile (10 mL) was added 2-(pyridin-3-yloxy)propane-1,3-diyl bis(4-methylbenzenesulfonate) (181-6, 330 mg, 0.690 mmol) and DIPEA (0.45 g, 3.5 mmol) and the resulting mixture was stirred at 120° C. for 16 h under microwave irradiation. The reaction mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted with DCM (3×30 mL). The combined organic phases were washed with brine, dried over anhydrous Na₂SO₄ and concentrated to dryness. The obtained crude material was purified by reverse phase HPLC (Column: ZORBAXECLIPSEXDB C18 (150 mm×19 mm), 5.0g, Mobile phase-A: 0.01% HCOOH (aq), Mobile phase-B: acetonitrile, Time (min)/% B: 0/5, 2/20, 10/40. Flow rate: 20 ml/min). The fractions containing the desired product were combined and lyophilized to afford I-203 (25.0 mg, 50.0 μmol, 9% yield) as white solid. MS [M+H]⁺=477.3. ¹H NMR (400 MHz, DMSO-d₆): δ 10.96 (s, 1H), 8.20-8.17 (m, 2H), 7.62 (d, J=8.4 Hz, 1H), 7.33-7.25 (m, 2H), 7.15 (s, 1H), 7.04 (d, J=8.4 Hz, 1H), 5.09 (dd, J=13.2, 4.8 Hz, 1H), 4.81-4.78 (m, 1H), 4.58-4.22 (m, 3H), 3.77-3.73 (m, 2H), 3.12-2.67 (m, 4H), 2.60-2.32 (m, 4H), 2.14-2.12 (m, 1H), 1.98-1.95 (m, 1H), 1.80-1.65 (m, 4H), 1.44-1.41 (m, 1H).

Example 182: 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-213) and 3-(5-(((1S,2S)-2-(diethylamino)cyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-214)

Step 1. 5-bromo-4-fluoro-3-hydroxyisobenzofuran-1(3H)-one (182-2)

To a stirred solution of TMP (9.7 mL, 57 mmol) in THF (40 mL) under an atmosphere of nitrogen was added BuLi (2.7 M in heptane, 20.3 mL, 54.7 mmol) dropwise at 0° C. and the resulting mixture was stirred for 30 min at 0° C. The reaction mixture was then cooled to about −45° C. (using dry ice/MeCN bath) and 4-bromo-3-fluorobenzoic acid (182-1, 4.99 g, 22.8 mmol), dissolved in THF (15 mL), was added dropwise and stirring was continued at −45° C. for 5 h. DMF (2.7 mL, 34 mmol) was then added dropwise and the reaction mixture was allowed to warm to rt and stirred overnight. The reaction mixture was quenched with aq. 3M HCl (40 mL) at 0° C. and extracted with DCM (×3). The combined organic phases were dried over Na₂SO₄, filtered, and concentrated to dryness. The crude product was purified via silica gel chromatography eluting with 0 to 100% EtOAc in heptane to afford 182-2 (2.91 g, 11.4 mmol, 50% yield) as a pale brown solid. MS [M+H]⁺=247.0. ¹H NMR (400 MHz, Acetonitrile-d₃) δ 7.90 (dd, J=8.0, 5.8 Hz, 1H), 7.56 (d, J=8.0 Hz, 1H), 6.72 (s, 1H), 5.92 (br s, 1H).

Step 2. 3-(5-bromo-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (182-3)

To a stirred solution of 182-2 (2.90 g, 11.7 mmol) in DMF (20 mL) was added 3-aminopiperidine-2,6-dione HCl salt (I-1c, 2.90 g, 17.6 mmol) and NaBH(OAc)₃ (6.22 g, 29.3 mmol) and the resulting mixture was stirred for 2 days at rt. The reaction mixture was diluted with H₂O (50 mL) and cooled to 0° C. with water/ice bath which resulted in the formation of precipitate. The resulting mixture was filtered and the dark blue solid was washed with Et₂O (×3). The obtained solid was dried in a vacuum oven to afford 182-3 (1.89 g, 5.31 mmol, 45% yield) as a grey solid. MS [M+H]⁺=341.1. ¹H NMR (400 MHz, DMSO-d₆) δ 11.02 (s, 1H), 7.88 (dd, J=8.0, 6.0 Hz, 1H), 7.55 (d, J=8.0 Hz, 1H), 5.12 (dd, J=13.3, 5.1 Hz, 1H), 4.62 (d, J=17.6 Hz, 1H), 4.45 (d, J=17.6 Hz, 1H), 2.99-2.85 (m, 1H), 2.66-2.55 (m, 1H), 2.47-2.36 (m, 1H), 2.05-1.96 (m, 1H).

Step 3. 3-(5-bromo-4-fluoro-1-oxoisoindolin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (182-4)

To a stirred suspension of 182-3 (500 mg, 1.47 mmol) and DBU (0.44 mL, 2.9 mmol) in DMF (4 mL) was added SEMCl (0.39 mL, 2.2 mmol) in one portion at 0° C. and the resulting mixture was stirred at room temperature overnight. The reaction mixture was quenched with sat. aq. NH₄Cl, extracted with EtOAc (×3). The combined organic phases were dried over sodium sulfate, filtered and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0 to 100% EtOAc in heptane to afford 182-4 (232 mg, 0.463 mmol, 32% yield) as an off-white solid. MS [M−H]⁻=469.1. ¹H NMR (400 MHz, Chloroform-d) δ 7.70 (dd, J=8.0, 5.8 Hz, 1H), 7.56 (d, J=8.1 Hz, 1H), 5.31-5.15 (m, 3H), 4.55 (d, J=16.4 Hz, 1H), 4.40 (d, J=16.4 Hz, 1H), 3.66-3.58 (m, 2H), 3.04 (ddd, J=17.9, 4.7, 2.5 Hz, 1H), 2.96-2.84 (m, 1H), 2.35 (qd, J=13.3, 4.7 Hz, 1H), 2.26-2.16 (m, 1H), 0.97-0.90 (m, 2H), 0.00 (s, 9H).

Step 4. tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-4-fluoro-1-oxoisoindolin-5-yl)oxy)cyclopentyl)carbamate (182-5)

To a mixture of 182-4 (232 mg, 0.492 mmol), tert-butyl ((1S,2S)-2-hydroxycyclopentyl)carbamate (1-1e, 149 mg, 0.738 mmol), NiCl₂(glyme) (5.4 mg, 0.025 mmol), dtbbpy (6.6 mg, 0.025 mmol) and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (5.5 mg, 4.9 μmol) in MeCN (4 mL) under nitrogen atmosphere was added 2,2,6,6-tetramethylpiperidine (TMP, 0.13 mL, 0.74 mmol). The resulting mixture was then stirred vigorously for 49 hours under irradiation of Blue LED lights at room temperature. The reaction mixture was filtered and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0 to 10% EtOH in DCM to afford 182-5 (207 mg, 0.175 mmol, purity ˜50%, 36% yield) as pale yellow solid. MS [M−H]⁻=590.1. The compound was carried forward to the next step without further purification.

Step 5. 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-213)

To a solution 182-5 (207 mg, 0.175 mmol, ˜50% purity) in MeCN (1 mL) was added methanesulfonic acid (0.11 mL, 1.7 mmol) and the resulting mixture was stirred overnight at rt. Et₃N (0.37 mL, 2.6 mmol) was then added dropwise at 0° C. and the reaction mixture was allowed to warm to rt. N,N-dimethylethylenediamine (0.03 mL, 0.3 mmol) was added and stirring was continued overnight at rt. The reaction mixture was diluted with sat. aq. NaHCO₃ and extracted with DCM:EtOH (v/v=4:1) (×3). The combined organic phases were concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM to afford I-213 (37 mg, 0.10 mmol, 58% yield) as white solid. MS [M+H]⁺=362.2. ¹H NMR (400 MHz, Acetonitrile-d₃) δ 7.50 (d, J=8.3 Hz, 1H), 7.27 (t, J=7.8 Hz, 1H), 5.03 (dd, J=13.4, 5.2 Hz, 1H), 4.53-4.27 (m, 3H), 3.38 (tt, J=5.8, 2.8 Hz, 1H), 2.88-2.66 (m, 2H), 2.41 (qd, J=13.2, 4.9 Hz, 1H), 2.23-2.15 (m, 1H), 2.13-2.07 (m, 1H, shoulder on residual H₂O peak), 2.04-1.97 (m, 3H), 1.82-1.68 (m, 3H), 1.46-1.34 (m, 1H).

Step 6. 3-(5-(((1S,2S)-2-(diethylamino)cyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-214)

To a stirred solution of I-213 (35 mg, 0.097 mmol) and NaBH(OAc)₃ (62 mg, 0.29 mmol) in DMF (1 mL) was added acetaldehyde (0.02 mL, 0.3 mmol) and the resulting mixture was stirred at rt for 35 min. The reaction mixture was concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM to afford I-214 (29 mg, 0.068 mmol, 70% yield) as white solid. MS [M+H]⁺=418.3. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 8.81 (s, 1H), 7.53 (d, J=8.3 Hz, 1H), 7.28 (ddd, J=8.5, 7.3, 2.3 Hz, 1H), 5.10 (dd, J=13.4, 5.2 Hz, 1H), 4.66 (dt, J=7.0, 3.1 Hz, 1H), 4.45-4.30 (m, 2H), 3.41-3.26 (m, 1H), 2.88-2.73 (m, 2H), 2.64-2.48 (m, 4H), 2.38-2.25 (m, 1H), 2.21-2.10 (m, 1H), 2.01-1.89 (m, 2H), 1.84-1.76 (m, 1H), 1.76-1.68 (m, 2H), 1.59-1.47 (m, 1H), 0.99 (t, J=7.1 Hz, 6H).

Example 183: 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-215) and 3-(5-(((1S,2S)-2-(diethylamino)cyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-216)

Step 1. Methyl 4-bromo-2-(bromomethyl)-5-fluorobenzoate (183-2)

To a stirred solution of 4-bromo-5-fluoro-2-methylbenzoate (183-1, 2700 mg, 10.93 mmol) in DCE (25 mL) under an atmosphere of nitrogen was added NBS (2140 mg, 12.02 mmol) followed by AIBN (90 mg, 0.55 mmol), and the resulting mixture was stirred vigorously at 85° C. for 8 h. The reaction mixture was quenched with sat. aq. Na₂S₂O₃ and then extracted with DCM (×3). The combined organic phases were concentrated to dryness. The crude product was purified via silica gel chromatography eluting with 0 to 50% EtOAc in heptane to afford 183-2 (3.37 g, 9.30 mmol, 85% yield) as a colorless oil. ¹H NMR (400 MHz, Chloroform-d) δ 7.73 (d, J=9.0 Hz, 1H), 7.69 (d, J=6.5 Hz, 1H), 4.89 (s, 2H), 3.95 (s, 3H).

Step 2. 3-(5-bromo-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (183-3)

To a solution of 183-2 (3.37 g, 9.30 mmol) in DMF (20 mL) was added 3-aminopiperidine-2,6-dione HCl salt (I-1c, 2.30 g, 14.0 mmol), followed by DIPEA (8.1 mL, 47 mmol), and the resulting mixture was stirred at 85° C. for 2 days. Excess DIPEA was removed by concentrating the mixture to a constant volume at 100 mbar and at a temperature of 40° C. The reaction mixture was then poured into conical flask containing H₂O (80 mL). The precipitate that formed was filtered and washed with H₂O (×2) and Et₂O (×2). The obtained solid was dried in the vacuum oven for 5 hours to afford 183-3 (2.22 g, 6.51 mmol, 70% yield) as a dark grey solid. MS [M+H]⁺=341.1. ¹H NMR (400 MHz, DMSO-d₆) δ 11.01 (s, 1H), 8.05 (d, J=6.0 Hz, 1H), 7.71 (d, J=7.7 Hz, 1H), 5.12 (dd, J=13.3, 5.1 Hz, 1H), 4.46 (d, J=17.5 Hz, 1H), 4.33 (d, J=17.5 Hz, 1H), 2.97-2.83 (m, 1H), 2.65-2.56 (m, 1H), 2.39 (qd, J=13.2, 4.5 Hz, 1H), 2.09-1.94 (m, 1H).

Step 3. 3-(5-bromo-6-fluoro-1-oxoisoindolin-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (183-4)

To a stirred solution of 183-3 (800 mg, 2.35 mmol) and DBU (0.70 mL, 4.7 mmol) in DMF (5 mL). was added SEMCl (0.62 mL, 3.5 mmol) at 0° C. and the resulting mixture was stirred at room temperature overnight. Additional DBU (0.70 mL, 4.7 mmol) and SEMCl (0.62 mL, 3.5 mmol) were added at 0° C. and stirring was continued for 2 hours at rt. The reaction mixture was quenched with sat. aq. NH₄Cl and extracted with EtOAc (×3). The combined organic phases were concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0 to 30% acetone in heptane to afford 183-4 (683 mg, 1.45 mmol, 62% yield) as a white solid. MS [M+H]⁺=471.2. ¹H NMR (400 MHz, Methylene Chloride-d) δ 7.73 (d, J=5.7 Hz, 1H), 7.57 (d, J=7.3 Hz, 1H), 5.23-5.09 (m, 3H), 4.36 (q, J=16.2 Hz, 2H), 3.62-3.57 (m, 2H), 2.99 (ddd, J=17.9, 4.8, 2.6 Hz, 1H), 2.87 (ddd, J=18.0, 13.3, 5.5 Hz, 1H), 2.32 (qd, J=13.2, 4.8 Hz, 1H), 2.25-2.14 (m, 1H), 0.94-0.87 (m, 2H), 0.00 (s, 9H).

Step 4. tert-butyl ((1S,2S)-2-((2-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-6-fluoro-1-oxoisoindolin-5-yl)oxy)cyclopentyl)carbamate (183-5)

To a mixture of 183-4 (683 mg, 1.45 mmol), tert-butyl ((1S,2S)-2-hydroxycyclopentyl)carbamate (1-1e, 350 mg, 1.74 mmol), NiCl₂(glyme) (16 mg, 0.072 mmol), dtbbpy (19 mg, 0.072 mmol) and Ir[(dF(CF₃)ppy)₂dtbbpy]PF₆ (16 mg, 0.014 mmol) in MeCN (8 mL) under an atmosphere of nitrogen was added 2,2,6,6-tetramethylpiperidine (0.37 mL, 2.2 mmol) and the resulting mixture was stirred vigorously for 48 hours under irradiation of Blue LED lights at room temperature. The reaction mixture was then filtered and concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0 to 100% EtOAc in heptane to afford 183-5 (420 mg, 0.639 mmol, 44% yield) as a yellow solid. MS [M+Na]⁺=614.2. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 7.62-7.49 (m, 1H), 7.47 (d, J=9.7 Hz, 1H), 5.22-5.04 (m, 3H), 4.78-4.67 (m, 1H), 4.57 (s, 1H), 4.40-4.25 (m, 2H), 4.12-3.99 (m, 1H), 3.63-3.56 (m, 2H), 3.02-2.79 (m, 2H), 2.40-2.24 (m, 1H), 2.22-2.13 (m, 2H), 2.12-2.02 (m, 1H), 1.96-1.85 (m, 2H), 1.84-1.71 (m, 1H), 1.60-1.49 (m, 1H), 1.42 (s, 9H), 0.94-0.87 (m, 2H), 0.00 (s, 9H).

Step 5. 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-215)

To a stirred solution of 183-5 (420 mg, 0.639 mmol) in MeCN (2 mL) was added methanesulfonic acid (0.5 mL, 7 mmol) and the resulting mixture stirred overnight at rt. Et₃N (1.5 mL, 11 mmol) was added dropwise at 0° C. and the reaction mixture was allowed to reach rt. N1,N2-dimethylethane-1,2-diamine (0.15 mL, 1.4 mmol) was then added and stirring was continued overnight at rt. The reaction mixture was diluted with sat. aq. NaHCO₃ and extracted with DCM:EtOH (v/v=4:1) (×4). The combined organic phases were concentrated to dryness. The crude product was purified by silica gel chromatography eluting with 0 to 100% EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM to afford I-215 (183 mg, 0.496 mmol, 78% yield) as white solid. MS [M+H]⁺=362.1. ¹H NMR (400 MHz, DMSO-d₆) δ 10.99 (s, 1H), 7.57 (d, J=9.8 Hz, 1H), 7.51 (t, J=6.0 Hz, 1H), 5.09 (dt, J=13.1, 4.6 Hz, 1H), 4.94-4.85 (m, 1H), 4.41 (dd, J=17.3, 7.2 Hz, 1H), 4.29 (dd, J=17.3, 8.0 Hz, 1H), 3.79-3.58 (m, 1H), 2.98-2.84 (m, 1H), 2.75-2.54 (m, 1H), 2.45-2.32 (m, 1H), 2.30-2.20 (m, 1H), 2.20-2.07 (m, 1H), 2.04-1.94 (m, 1H), 1.87-1.61 (m, 4H).

Step 6. 3-(5-(((1S,2S)-2-(diethylamino)cyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (I-216)

To a stirred solution of I-215 (150 mg, 0.415 mmol) and NaBH(OAc)₃ (264 mg, 1.25 mmol) in DMF (2 mL) was added acetaldehyde (0.07 mL, 1 mmol) and the resulting mixture was stirred at rt overnight. The reaction mixture was concentrated to dryness. The crude product was purified by silica gel chromatography eluting with EtOAc:EtOH (v/v=3:1, with 1% Et₃N) in DCM to afford I-216 (87.5 mg, 0.206 mmol, 50% yield) as white solid. MS [M+H]⁺=418.2. ¹H NMR (400 MHz, Methylene Chloride-d₂) δ 8.45 (br s, 1H), 7.51 (dd, J=9.8, 3.1 Hz, 1H), 7.37 (t, J=7.3 Hz, 1H), 5.16 (ddd, J=13.3, 5.3, 3.5 Hz, 1H), 4.74 (s, 1H), 4.48-4.17 (m, 2H), 3.51-3.34 (m, 1H), 2.96-2.80 (m, 2H), 2.73-2.56 (m, 4H), 2.45-2.28 (m, 1H), 2.27-2.17 (m, 1H), 2.10-1.96 (m, 2H), 1.93-1.74 (m, 3H), 1.64 (s, 1H), 1.07 (t, J=7.1 Hz, 6H).

Biological Assays and Data

The activity of a compound according to the present disclosure can be assessed by the following in vitro methods.

Example 184: Prolabel Quantification of IKZF1, IKZF2, or GSPT1 Protein Levels in 293GT Cells

The Prolabel system from DiscoverX was used to develop high-throughput and quantitative assays to measure changes in IKZF1, IKZF2 and GSPT1 protein levels in response to compounds. The prolabel tag is derived from the alpha fragment of beta galactosidase and has the following protein sequence: mssnslavvlqrrdwenpgvtqlnrlaahppfaswrnseeartdrpsqqlrslnge. The complementary fragment of beta-galactosidase (from DiscoverX), is added to the prolabel tag to form an active beta galactosidase enzyme whose activity can be precisely measured. In this way, the levels of a fusion protein with the prolabel tag can be quantified in cell lysates.

Lentiviral vectors, based on the Invitrogen pLenti6.2/V5 DEST backbone, were constructed that placed the prolabel tag upstream of IKZF1, IKZF2 or GSPT1 and expressed the fusion protein from a CMV promoter.

To ensure moderate and consistent expression of the prolabel fusion proteins across all cells in the population, stable cell lines were constructed from cells expressing a single copy of the construct. Lentivirus packaged with the constructs was made using the Virapower kit from Invitrogen. Strongly adherent 293GT cell, GripTite 293 MSR cells from Thermo Fisher Scientific (Catalog number: R79507), were infected with the virus at low multiplicity of infection and selected by 5 μg/mL blasticidin for 2 weeks.

The levels of prolabel tagged fusion proteins in compound treated cell lines were measured as follows:

Day 1, Cells were diluted to 1.0×10⁶ cells/ml in normal growth medium. 17.5 μL of cells were plated in each well of a solid white 384 well plate. Plates were incubated overnight in a 37° C. tissue culture incubator.

Day 2, Serial dilutions of compounds were made in 384 well plates from 10 mM stocks. 15 μL of DMSO was added to each well of a 384 well plate. In the first column 15 μL of stock compound was added. The solution was mixed and 15 μL was transferred to the next column. This was repeated until 20 two-fold dilutions were prepared. 2.5 μL of diluted compounds were transferred into 60 μL of cell culture medium in another 384 well plate, and mixed well. 2.5 μL of this mixture was added to the plated cells. The final DMSO concentration was 0.5% and the highest concentration of compound was 50 μM. Plates were incubated overnight (e.g., about 14 h, 18 h, or 24 h) in a 37° C. tissue culture incubator.

Day 3, Plates were removed from the incubator and allowed to equilibrate at rt for 30 minutes. Prolabel substrate (DiscoverX PathHunter Prolabel Detection Kit, User manual: 93-0180) was added as described by the manufacturers protocols. Plates were incubated at rt for three hours and luminescence was read using an Envision reader (Perkin Elmer) Data was analyzed and visualized using the Spotfire software package.

Table 14 shows Helios (IKZF2), Ikaros (IKZF1) and G1 to S phase transition 1 protein (GSPT1) degradation activity of compounds of the disclosure in Pro-label assays in 293GT cells, (% degradation is at 10 μM).

IKZF2% protein IKZF2 reduction IKZF1 GSPT1 Cmpd EC₅₀ at 10 μM, EC₅₀ EC₅₀ No. (mM) 24 h (mM) (mM) I-1 — 10% >30 >30 I-2 — 10% >30 >30 I-3 0.256 40% >30 — I-4 4.2 25% >30 — I-5 0.170 60% >30 — I-6 —  5% >30 — I-7 0.037 85% >30 — I-8 0.176 50% >30 >30 I-9 >30  0% >30 — I-10 >30  0% >30 — I-11 0.129 75% >30 >30 I-12 0.139 65% >30 >30 I-13 0.016 85% >30 >30 I-14 0.282 60% >30 >30 I-15 0.565 50% >30 >50 I-16 0.032 85% >30 >30 I-17 0.018 80% >30 — I-18 0.266 50% >30 — I-19 — 10% >30 — I-20 0.025 75% >30 >30 I-21 0.237 30% >30 >30 I-22 0.016 80% >50 >30 I-23 0.067 80% >30 >30 I-24 0.064 80% >30 >30 I-25 0.025 80% >30 >30 I-26 >30  0% >30 — I-27 0.548 50% >30 — I-28 >30  0% >30 >30 I-29 0.028 80% >30 — I-30 >30  0% >30 >30 I-31 0.006 90%    3.2 >30 I-32 0.011 85% >30 >30 I-33 0.163 55% >30 >30 I-34 >30  0% >50 — I-35 — 10% >30 >30 I-36 0.037 80% >30 — I-37 >30  0% >50 >50 I-49 0.007 86% >30 >30 I-50 0.007 85% >30 — I-51 0.006 90% >30 >30 I-52 0.005 89% >30 — I-53 0.003 88% >30 — I-48 0.003 89% >30 >30 I-54 0.001 91% >30 — I-55 0.004 91% >30 >30 I-56 0.002 91% >30 >30 I-38 0.015 86% >30 >50 I-39 0.11 57% >30 — I-43 >30  0% >30 — I-44 0.465 42% >30 — I-46 >30  0% >30 — I-47 >30 11% >30 — I-136 0.131 58% >30 — I-96 0.099 81% >30 — I-137 0.030 80% >30 — I-138 0.049 83% >30 — I-139 0.048 83% >30 — I-141 0.157 77% >30 — I-142 0.034 83% >30 — I-143 0.022 81% >30 — I-144 0.015 88% >30 — I-145 0.161 52% >30 — I-146 0.014 83% >30 — I-147 0.159 44% >30 — I-148 9.938 33% >30 — I-149 0.012 77% >30 — I-150 0.243 51% >30 — I-151 0.443 53% >30 — I-152 0.052 82% >30 — I-153 0.014 85% >30 — I-154 0.034 83% >30 — I-155 0.485 65%    0.10 — I-156 0.133 70% >30 — I-157 0.071 61% >30 — I-158 0.027 82% >30 — I-159 0.040 75% >30 >30 I-160 0.077 75% >30 — I-161 0.032 80% >30 — I-162 0.082 64% >30 — I-163 0.039 78% >30 — I-164 0.23 71% >30 — I-165 0.071 72% >30 — I-166 0.007 81% >30 — I-167 0.062 75% >30 — I-168 0.009 83% >30 — I-169 >30  2% >30 — I-170 0.194 67% >30 — I-171 0.021 74% >30 — I-172 0.124 58% >30 — I-173 0.037 84% >30 >30 I-174 0.085 86% >30 — I-175 — — >30 — I-176 0.027 89% >30 — I-177 0.002 92% >30 — I-178 0.006 92% >30 — I-179 0.007 85% >30 — I-180 0.017 81% >30 — I-182 0.017 76% >30 — I-183 0.010 84% >30 — I-184 0.053 38% >30 — I-185 0.096 66% — — I-187 0.012 79% >30 — I-186 0.007 86% >30 — I-188 0.003 70% >30 — I-189 0.104 70% >30 — I-190 1.089 39% >30 — I-191 0.065 83% >30 >30 I-192 0.035 77% >30 — I-193 0.039 79% >30 — I-194 0.015 83% >30 >30 I-195 0.005 85% >30 >30 I-196 0.084 68% >30 — I-197 0.084 68% >30 — I-198 0.076 67% >30 — I-199 0.130 71% >30 — I-200 0.057 71% >30 — I-201 0.260 46% >30 — I-71 0.018 88% >30 — I-72 0.361 57% >30 — I-73 0.136 66% >30 — I-74 0.037 58% >30 — I-75 0.065 79% >30 — I-76 0.137 79% >30 — I-77 0.141 86% >30 — I-78 0.880 50% >30 — I-80 0.263 65% >30 — I-81 0.046 85% >30 — I-82 2.64 58% >30 — I-83 0.029 86% >30 — I-84 0.118 35% >30 — I-85 0.087 69% >30 — I-86 0.177 76% >30 — I-87 0.082 79% >30 — I-88 5.528 57% >30 — I-89 0.038 82% >30 >30 I-90 0.246 57% >30 — I-91 0.129 58% >30 — I-92 0.124 60% >30 — I-93 0.345 60% >30 — I-95 0.036 84% >30 — I-94 0.074 84% >30 — I-97 0.592 36% >30 — I-98 0.055 80% >30 — I-101 0.067 73% >30 — I-102 0.067 73% >30 — I-103 0.060 54% — — I-104 0.028 87% >30 — I-105 0.234 52% >30 — I-106 0.411 51% >30 — I-107 0.016 89% >30 — I-110 0.022 89% >30 — I-111 0.364 59% >30 — I-112 0.0002 95% >30 — I-113 0.0008 93% >30 >30 I-114 0.458 54% >30 — I-115 0.008 90% >30 — I-116 0.041 65% >30 — I-119 0.091 59% — — I-118 0.045 67% — — I-120 0.212 66% >30 >30 I-121 0.0003 94% >30 — I-122 0.027 84% >30 >30 I-123 0.196 82% >30 — I-129 0.082 70% >30 — I-130 0.050 50% >30 — I-79 0.027 85% >30 — I-131 0.012 91% >30 — I-132 0.134 80% >30 — I-133 0.045 81% >30 — I-134 0.207 56% >30 — I-135 0.003 92% — >30 I-57 0.047 80% >30 — I-58 0.064 75% >30 — I-59 1.31 39% >30 — I-60 0.157 61% >30 — I-61 0.028 84% >30 >30 I-62 0.057 88 >30 — I-63 0.011 85% >30 >30 I-64 0.036 84% >30 — I-65 0.157 76% >30 — I-66 0.096 81% >30 — I-67 0.066 74% >30 — I-68 0.004 90% >30 >30 I-69 0.014 84% >30 >30 I-99 0.11 86% >30 — I-100 0.044 86% >30 — I-213 >30 20% >30 — I-214 0.33 40% >30 — I-215 >30 30% >30 — I-216 0.12 60% >30 —

Example 185: Quantification of In Vitro Suppressive Potency of Primary Human Regulatory T Cells Expanded in the Presence of Compounds

Materials and Methods

Treg Cell Sorting:

Human buffy coats are obtained from BioreclamationIVT, in the USA. CD4+ T cells are isolated from said buffy coats using the RosetteSep Human CD4+ T cell enrichment Cocktail (Stemcell technologies, USA) and gradient centrifugation over Ficoll Paque Plus (GE HealthCare LifeSciences, USA) as per manufacturer's recommendations. Cells are resuspended in RPMI medium supplemented with 1% penicillin-Streptomycin solution, 10% Fetal Bovine Serum, HEPES (10 mM), MEM NEAA (100 nM), sodium pyruvate (1 mM) (all supplements from Thermo Fisher Scientific, USA), thereafter referred to as complete RPMI (cRPMI), and rested overnight at 37° C., 5% CO₂ in the presence of 2U/mL rhIL-2 (Proleukin, Novartis). Cells are collected and resuspended in autoMACS Running Buffer supplemented with BSA (Miltenyi Biotec, USA) and labelled using CD4-FITC antibody (clone RPA-T4), CD25-APC antibody (clone M-A251) (Biolegend) and CD25 Microbeads (Miltenyi Biotec, USA). CD25-enriched cells are then isolated using the autoMACS Pro Separator. A highly purified population of Treg cells are then obtained by further sorting CD4+ CD25Hi cells using a Sony SH800 cell sorter. The resulting Treg cell population is routinely above 90% pure according to FOXP3 expression.

Treg Cell Expansion:

Purified Treg cells are plated in cRPMI in 96-well, round-bottom plates at a density of 25000-50000 cells per well and activated in the presence of 500 U/mL rhIL2, and Treg expander Dynabeads (Thermo Fisher Scientific, USA) according to manufacturer's recommendations, in the presence or absence of 100 μM rapamycin (Thermo Fisher Scientific, USA). The compounds of the present disclosure are then added at a final concentration of 10 M and DMSO is added as a vehicle control. Cells are incubated at 37° C., 5% CO₂ for a total of 12-14 days. The compound and rhIL2 are replenished every 48h during the entirety of the culture.

Phenotypic Analysis of Expanded Treg Cells:

Cell are collected and counted and the fold expansion is calculated as (number of cells recovered)/(number of cells plated). A fraction of the cells is fixed and permeabilized using the eBioscience Foxp3 staining Buffer kit (eBioscience, Thermo Fisher Scientific, USA) and stained with Helios-PECyanine7 antibody (Clone 22F6). To determine IL2-expression, expanded Treg cells are further incubated in the presence of the eBioscience Cell Stimulation Cocktail with Protein inhibitors (Thermo Fisher Scientific) for 4 hours, followed by fixation and staining with IL2-BV711 antibody (clone MQ1-17H12) (Biolegend, USA). Cells are acquired on an LSRFortessa (Becton Dickinson, USA) and analysis is performed using the FlowJo software (TreeStar, USA).

Functional Analysis of Expanded Treg Cells:

Primary human PBMCs are obtained from freshly prepared buffy coats (BioReclamationIVT) using gradient centrifugation over Ficoll Paque Plus as per manufacturer's recommendations. Cells are then labelled with CFSE (5(6)-Carboxyfluorescein diacetate N-succinimidyl ester, Sigma-Aldrich, USA) and plated in triplicates cRPMI in round bottom 96-well plates, alone or with expanded Treg cells at a 1:2 PBMC:Treg ratio. The compounds of the present disclosure are then added at a final concentration of 10 M and DMSO is added as a vehicle control. Cells are activated using soluble anti-CD3 antibody (clone OKT3) (eBioscience, ThermoFisher Scientific, USA) at a final concentration of 100 ng/ml. Cells are incubated at 37° C., 5% CO₂ for a total of 4-5 days. At the end of the culture, cells are stained using the Live/dead Blue viability stain (Thermo Fisher Scientific, USA) as per manufacturer's instructions, followed by staining with CD4-BUV737 (Clone SK3) (BDBiosciences, USA) and CD8-BV711 (clone RPA-T8) (Biolegend, USA). Cells are acquired on an LSRFortessa (Becton Dickinson, USA) and analysis is performed using the FlowJo software (TreeStar, USA). Proliferation is assessed in each population as the proportion of cells having diluted CFSE. Suppression is assessed for each condition in comparison to the responders plated alone.

Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments described specifically herein. Such equivalents are intended to be encompassed in the scope of the following claims. 

What is claimed is:
 1. A compound of Formula (I′)

wherein: X₁ and X₂ are each independently H, (C₁-C₄)alkyl, (C₁-C₆)alkoxy, (C₁-C₄)haloalkyl, (C₁-C₆)haloalkoxy, (C₃-C₇)cycloalkyl, halogen, —CN, —OH, or —NH₂, R_(x) is H or D; R₁ is

each R₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —CN, —OH, or —NH₂; or two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring consisting of 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring consisting of 1-3 heteroatoms selected from O, N, and S; or R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring optionally consisting of 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂; each R₃ is (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —OH, or —NH₂; R₄ is —OR₅ or —NR₆R_(6′); R₅ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S; R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₂-C₆)hydroxyalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇ and wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₁₂; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 8-membered heterocycloalkyl ring optionally consisting of 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈; or R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 4- to 6-membered heterocycloalkyl ring optionally consisting of 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂; each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₉; each R₈ is independently at each occurrence halogen, (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, —CN, —OH, —NR₁₃R₁₄, —NH₂, —O(C₃-C₇)cycloalkyl, —O-4- to 7-membered heterocycloalkyl ring consisting of 1-3 heteroatoms selected from O, N, and S, —O(C₆-C₁₀)aryl, or —O-5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S, wherein the alkoxy is optionally substituted with one to three R₁₀ and the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to three R₁₁; or two R₈ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring consisting of 1-2 heteroatoms selected from O, N, and S optionally substituted with two R₁₅; or two R₈ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S; or two R₈ together with the same atom to which they are attached form a ═(O); each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, (C₃-C₆)cycloalkyl, —OH, —CN, —NH₂, or —NR₁₃R₁₄; or two R₉ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring consisting of 1-2 heteroatoms selected from O, N, and S optionally substituted with one or more substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂; or two R₉ when on adjacent atoms together with the atoms to which they are attached form a (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S; each R₁₀ is independently at each occurrence selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S; each R₁₁ is independently at each occurrence selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, and —NH₂; each R₁₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkoxy, halogen, —OH, —CN, or —NH₂; two R₁₂ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring consisting of 1-2 heteroatoms selected from O, N, and S; R₁₃ and R₁₄ are each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S; two R₁₅ together with the atoms to which they are attached form a (C₄-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring consisting of 1-2 heteroatoms selected from O, N, and S; m and m1 are each independently 0, 1 or 2; n1 is 0, 1, 2, or 3; and each s and n is independently 1, 2, or 3, wherein s+n is ≤4; or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 2. The compound according to claim 1, having a Formula (I),

wherein: R_(x) is H or D; R₁ is

each R₂ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —CN, —OH, or —NH₂; or two R₂ together with the carbon atoms to which they are attached form a (C₃-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring consisting of 1-3 heteroatoms selected from O, N, and S; or two R₂ together when on adjacent carbon atoms form a phenyl or a 5- or 6-membered heteroaryl ring consisting of 1-3 heteroatoms selected from O, N, and S; or R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally consisting of 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂; each R₃ is (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, halogen, —OH, or —NH₂; R₄ is —OR₅ or —NR₆R_(6′); R₅ is H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three substituents independently selected from (C₆-C₁₀)aryl and 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S; R₆ and R_(6′) are each independently H, (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, (C₃-C₇)cycloalkyl, 5- or 6-membered heterocycloalkyl consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S, wherein the alkyl is optionally substituted with one to three R₇; or R₆ and R_(6′) together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycloalkyl ring optionally consisting of 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four R₈; or R₂ and R₆ together with the carbon and nitrogen atoms to which they are attached form a 5- or 6-membered heterocycloalkyl ring optionally consisting of 1-2 additional heteroatoms selected from O, N, and S, and optionally substituted with one to four substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂; each R₇ is (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, or 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S, wherein the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with one to four R₉; each R₃ is independently at each occurrence halogen, (C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₁-C₆)haloalkyl, (C₁-C₆)haloalkoxy, —CN, —OH, or —NH₂, wherein the alkoxy is optionally substituted with one to three substituents independently selected from (C₃-C₇)cycloalkyl, 4- to 7-membered heterocycloalkyl ring consisting of 1-3 heteroatoms selected from O, N, and S, (C₆-C₁₀)aryl, and 5- or 6-membered heteroaryl consisting of 1-3 heteroatoms selected from O, N, and S; or two R₈ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 4- to 7-membered heterocycloalkyl ring consisting of 1-2 heteroatoms selected from O, N, and S; each R₉ is independently at each occurrence (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, or —NH₂; or two R₉ together with the atoms to which they are attached form a (C₅-C₇)cycloalkyl or a 5- to 7-membered heterocycloalkyl ring consisting of 1-2 heteroatoms selected from O, N, and S optionally substituted with one or more substituents each independently selected from (C₁-C₆)alkyl, (C₁-C₆)haloalkyl, halogen, —OH, —CN, and —NH₂; m and m1 are each independently 0, 1, or 2; n1 is 0, 1, 2, or 3; and each s and n is independently 1, 2, or 3, wherein s+n is ≤4; or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 3. The compound according to claim 1, wherein R₁ is


4. The compound according to claim 1, wherein n1 is 0, 1, or
 2. 5. The compound according to claim 1, wherein m1 is
 0. 6. The compound according to claim 1, wherein m1 is
 2. 7. The compound according to claim 1, wherein R₁ is


8. The compound according to claim 7, wherein n is 2 and s is 1 or
 2. 9. The compound according to claim 8, wherein m is 0 or
 1. 10. The compound according to claim 1, having a Formula (Ia), Formula (Ib), Formula (Ic), Formula (Id), Formula (Ie), Formula (ID, Formula (Ig), Formula (Ih), Formula (Ii), Formula (Ij), Formula (Ik), Formula (II), Formula (Im), Formula (In), Formula (Io), or Formula (Ip):

or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 11. The compound according to claim 10, wherein R₄ is —OR₅.
 12. The compound according to claim 10, wherein R₄ is —NR₆R_(6′).
 13. The compound according to claim 1, having a Formula (Iq), Formula (Ir), Formula (Is), or Formula (It):

or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 14. The compound according to claim 1 selected from: 3-(5-(((1S,2S)-2-((2,2-difluoroethyl)(ethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((2,2-difluoroethyl)(ethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((2,2-difluoroethyl)(ethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((2,2-difluoroethyl)(ethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((2,2-difluoroethyl)(ethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((1-methyloctahydrocyclopenta[b]pyrrol-6-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-methoxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-methoxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-methoxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-methoxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-methoxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-hydroxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-hydroxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-hydroxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-hydroxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-hydroxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-hydroxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-hydroxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-hydroxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-hydroxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-hydroxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(isobutylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(isobutylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(isobutylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(isobutylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(isobutylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4,4-difluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4,4-difluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4,4-difluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4,4-difluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4,4-difluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(benzyloxy)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(benzyloxy)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(benzyloxy)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(benzyloxy)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(benzyloxy)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(diethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(((1-(trifluoromethyl)cyclopropyl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(((1-(trifluoromethyl)cyclopropyl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(((1-(trifluoromethyl)cyclopropyl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(((1-(trifluoromethyl)cyclopropyl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(((1-(trifluoromethyl)cyclopropyl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-aminocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-aminocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-phenoxycyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(benzylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(benzylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(benzylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(benzylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(benzylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3S,4S)-3-(benzylamino)tetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3R,4S)-3-(benzylamino)tetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3R,4R)-3-(benzylamino)tetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3S,4R)-3-(benzylamino)tetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((3-(benzylamino)tetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(benzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(((R)-1-phenylethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethyl(2-fluoroethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl(isopropyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethyl(isopropyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethyl(isopropyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethyl(isopropyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethyl(isopropyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-methoxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-methoxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-methoxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-methoxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-methoxycyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-hydroxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-hydroxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-hydroxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-hydroxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-hydroxycyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(bis(cyclopropylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(bis(cyclopropylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(bis(cyclopropylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(bis(cyclopropylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(bis(cyclopropylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(piperidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(piperidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(piperidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(piperidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(piperidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-morpholinocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-morpholinocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-morpholinocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-morpholinocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-morpholinocyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(dibenzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(dibenzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(dibenzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(dibenzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(dibenzylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; cis-3-(5-((2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; trans-3-(5-((2-(diethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(methylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(methylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(methylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(methylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(methylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 1-((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-methylpiperidine-4-carbonitrile; 1-((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-methylpiperidine-4-carbonitrile; 1-((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-methylpiperidine-4-carbonitrile; 1-((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-methylpiperidine-4-carbonitrile; 1-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-methylpiperidine-4-carbonitrile; 3-(5-(((1S,2S)-2-(benzyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(benzyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(benzyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(benzyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(benzyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3S,4S)-3-aminotetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3R,4S)-3-aminotetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3R,4R)-3-aminotetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3S,4R)-3-aminotetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((3-aminotetrahydro-2H-pyran-4-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((1S,2S)-2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((1R,2R)-2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((1S,2R)-2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((1R,2S)-2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(2-(benzylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3R,4S)-4-aminotetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3R,4R)-4-aminotetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3S,4R)-4-aminotetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3S,4S)-4-aminotetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((4-aminotetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3R,4S)-4-(diethylamino)tetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3R,4R)-4-(diethylamino)tetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3S,4R)-4-(diethylamino)tetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((3S,4S)-4-(diethylamino)tetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((4-(diethylamino)tetrahydrofuran-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 3-(5-((1S,2S)-2-(diethylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((1R,2S)-2-(diethylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((1R,2R)-2-(diethylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((1S,2R)-2-(diethylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(2-(diethylamino)cyclobutoxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethyl((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethyl((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethyl((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethyl((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 3-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 3-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 3-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 3-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 3-(5-(((1S,2S)-2-(isopropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(isopropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(isopropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(isopropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(isopropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 2-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 2-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 2-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 2-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 2-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 3-(5-(((1S,2S)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-hydroxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-hydroxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-hydroxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-hydroxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-hydroxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-hydroxy-4-(trifluoromethyl)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-hydroxy-4-(trifluoromethyl)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-hydroxy-4-(trifluoromethyl)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-hydroxy-4-(trifluoromethyl)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-hydroxy-4-(trifluoromethyl)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 1-((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-(trifluoromethyl)piperidine-4-carbonitrile; 1-((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-(trifluoromethyl)piperidine-4-carbonitrile; 1-((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-(trifluoromethyl)piperidine-4-carbonitrile; 1-((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-(trifluoromethyl)piperidine-4-carbonitrile; 1-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)-4-(trifluoromethyl)piperidine-4-carbonitrile; 3-(5-(((1S,2S)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-(2,2-difluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(2,2-difluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(2,2-difluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(2,2-difluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(2,2-difluoroethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-(cyclopropylmethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(cyclopropylmethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(cyclopropylmethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(cyclopropylmethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(cyclopropylmethoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-(benzyloxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(benzyloxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(benzyloxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(benzyloxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(benzyloxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-isopropoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-isopropoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-isopropoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-isopropoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-isopropoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-ethoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-ethoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-ethoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-ethoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-ethoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-(benzyloxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(benzyloxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(benzyloxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(benzyloxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(benzyloxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-ethoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-ethoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-ethoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-ethoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-ethoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-(3,3-difluorocyclobutoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(3,3-difluorocyclobutoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(3,3-difluorocyclobutoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(3,3-difluorocyclobutoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(3,3-difluorocyclobutoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-hydroxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(4-oxopiperidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-hydroxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-hydroxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-hydroxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-hydroxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-hydroxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-hydroxy-3-methylazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-hydroxy-3-methylazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-hydroxy-3-methylazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-hydroxy-3-methylazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-hydroxy-3-methylazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(isobutylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(isobutylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(isobutylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(isobutylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(isobutylamino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl(methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethyl(methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethyl(methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethyl(methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethyl(methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 3-(1-oxo-5-(((1S,2S)-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((3aR,6aS)-tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(tetrahydro-1H-furo[3,4-c]pyrrol-5(3H)-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((pyridin-2-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((pyridin-2-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((pyridin-2-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione 3-(1-oxo-5-(((1R,2S)-2-((pyridin-2-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((pyridin-2-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(pyrrolidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(pyrrolidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(pyrrolidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(pyrrolidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(pyrrolidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(bis((3-methyloxetan-3-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(bis((3-methyloxetan-3-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(bis((3-methyloxetan-3-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(bis((3-methyloxetan-3-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(bis((3-methyloxetan-3-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((pyridin-3-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((pyridin-3-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((pyridin-3-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((pyridin-3-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((pyridin-3-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((pyridin-4-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((pyridin-4-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((pyridin-4-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((pyridin-4-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((pyridin-4-ylmethyl)amino)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-methoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4,4-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4,4-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4,4-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4,4-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4,4-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-methoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-methoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-methoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-methoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-methoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl(oxetan-3-ylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethyl(oxetan-3-ylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethyl(oxetan-3-ylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethyl(oxetan-3-ylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethyl(oxetan-3-ylmethyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(isoindolin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(isoindolin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(isoindolin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(isoindolin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(isoindolin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-methoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-methoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-methoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-methoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-methoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-ethoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-ethoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-ethoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-ethoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-ethoxy-4-methylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((((1R,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((((1R,4R)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((((1R,4R)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((((1R,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((((1R,4R)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; (1S,4R)-4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)amino)methyl)cyclohexane-1-carbonitrile; (1R,4r)-4-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)amino)methyl)cyclohexane-1-carbonitrile; (1R,4r)-4-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)amino)methyl)cyclohexane-1-carbonitrile; (1S,4r)-4-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)amino)methyl)cyclohexane-1-carbonitrile; (1r,4r)-4-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)amino)methyl)cyclohexane-1-carbonitrile; 4-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)amino)methyl)cyclohexane-1-carbonitrile; 3-(5-(((1S,2S)-2-(((4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1S,3R)-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1R,3S)-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; trans-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; cis-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclopentyl)(ethyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; 3-(5-(((1S,2S)-2-(4-fluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-fluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-fluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-fluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-fluoropiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(1,5-oxazocan-5-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(1,5-oxazocan-5-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(1,5-oxazocan-5-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(1,5-oxazocan-5-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(1,5-oxazocan-5-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl(((1R,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((((1R,4S)-4-methoxycyclohexyl)methyl)(methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; trans-3-(5-((2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; cis-3-(5-((2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((4,4-difluorocyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((4,4-difluorocyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((4,4-difluorocyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((4,4-difluorocyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((4,4-difluorocyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((1H-indol-5-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((1H-indol-5-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((1H-indol-5-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((1H-indol-5-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((1H-indol-5-yl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-(tert-butoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-(tert-butoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-(tert-butoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-(tert-butoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-(tert-butoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(2-oxa-8-azaspiro[4.5]decan-8-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(6-azaspiro[3.5]nonan-6-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl(((1S,3R)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethyl(((1s,3S)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethyl(((1s,3S)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethyl(((1s,3R)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethyl(((1s,3s)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(diethylamino)-4,4-dimethylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl(((1R,3S)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethyl(((1r,3R)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethyl(((1r,3R)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethyl(((1r,3S)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethyl(((1r,3r)-3-methoxycyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(8-oxa-2-azaspiro[4.5]decan-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-thiomorpholinocyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-thiomorpholinocyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-thiomorpholinocyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-thiomorpholinocyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-thiomorpholinocyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(1,4-oxazepan-4-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(1,4-oxazepan-4-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(1,4-oxazepan-4-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(1,4-oxazepan-4-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(1,4-oxazepan-4-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-isopropoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-isopropoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-isopropoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-isopropoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-isopropoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-(cyclopropylmethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-(cyclopropylmethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-(cyclopropylmethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-(cyclopropylmethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-(cyclopropylmethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((3aR,4R,7S,7aS)-octahydro-2H-4,7-epoxyisoindol-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((3aR,4R,7S,7aS)-octahydro-2H-4,7-epoxyisoindol-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((3aR,4R,7S,7aS)-octahydro-2H-4,7-epoxyisoindol-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((3aR,4R,7S,7aS)-octahydro-2H-4,7-epoxyisoindol-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((3aR,4R,7S,7aS)-octahydro-2H-4,7-epoxyisoindol-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-ethoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-ethoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-ethoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-ethoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-ethoxypiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(7,8-dihydro-1,6-naphthyridin-6(5H)-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(7,8-dihydro-1,6-naphthyridin-6(5H)-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(7,8-dihydro-1,6-naphthyridin-6(5H)-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(7,8-dihydro-1,6-naphthyridin-6(5H)-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(7,8-dihydro-1,6-naphthyridin-6(5H)-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((((1S,4R)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((((1s,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((((1s,4S)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((((1s,4R)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((((1s,4s)-4-methoxycyclohexyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((R)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((S)-3-methoxypyrrolidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3,3-difluorocyclobutyl)methyl)amino)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-(difluoromethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-(difluoromethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-(difluoromethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-(difluoromethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-(difluoromethoxy)piperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(1,3-dihydro-2H-pyrrolo[3,4-c]pyridin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(1,3-dihydro-2H-pyrrolo[3,4-c]pyridin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(1,3-dihydro-2H-pyrrolo[3,4-c]pyridin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(1,3-dihydro-2H-pyrrolo[3,4-c]pyridin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(1,3-dihydro-2H-pyrrolo[3,4-c]pyridin-2-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(propylamino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(propylamino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(propylamino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(propylamino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(propylamino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(dipropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(dipropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(dipropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(dipropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(dipropylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((pyridin-4-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((pyridin-4-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((pyridin-4-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((pyridin-4-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((pyridin-4-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((pyridin-3-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((pyridin-3-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((pyridin-3-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((pyridin-3-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((pyridin-3-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((1-ethyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((1-ethyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((1-ethyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((1-ethyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((1-ethyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((1-isopropyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((1-isopropyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((1-isopropyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((1-isopropyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((1-isopropyl-1H-pyrazol-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethyl(methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(dimethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(dimethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(dimethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(dimethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(dimethylamino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((oxetan-3-ylmethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((oxetan-3-ylmethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((oxetan-3-ylmethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((oxetan-3-ylmethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((oxetan-3-ylmethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((2-hydroxyethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((2-hydroxyethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((2-hydroxyethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((2-hydroxyethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((2-hydroxyethyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(pyrrolidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(pyrrolidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(pyrrolidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(pyrrolidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(pyrrolidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-morpholinocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-morpholinocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-morpholinocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-morpholinocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-morpholinocyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((pyridin-2-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((pyridin-2-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((pyridin-2-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((pyridin-2-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((pyridin-2-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((3-hydroxy-3-methylbutyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((3-hydroxy-3-methylbutyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((3-hydroxy-3-methylbutyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((3-hydroxy-3-methylbutyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((3-hydroxy-3-methylbutyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((3-methyloxetan-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3-methyloxetan-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3-methyloxetan-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3-methyloxetan-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3-methyloxetan-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-methoxy-4-methylpiperidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-methoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-methoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-methoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-methoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-methoxyazetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((6-methylpyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((6-methylpyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((6-methylpyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((6-methylpyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((6-methylpyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((5-methoxypyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((5-methoxypyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((5-methoxypyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((5-methoxypyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((5-methoxypyridin-2-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((6-methoxypyridin-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((6-methoxypyridin-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((6-methoxypyridin-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((6-methoxypyridin-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((6-methoxypyridin-3-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((2-hydroxy-2-methylpropyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((2-hydroxy-2-methylpropyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((2-hydroxy-2-methylpropyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((2-hydroxy-2-methylpropyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((2-hydroxy-2-methylpropyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; 3-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; 3-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; 3-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; 3-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1S,3R)-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1R,3R)-3-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1R,3R)-3-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1S,3r)-3-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1r,3r)-3-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1R,3S)-3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1S,3s)-3-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1S,3s)-3-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1R,3s)-3-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; (1s,3s)-3-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)-1-methylcyclobutane-1-carbonitrile; 3-(1-oxo-5-(((1S,2S)-2-(piperidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(piperidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(piperidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(piperidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(piperidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((((1S,3R)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((((1s,3S)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((((1s,3S)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((((1s,3R)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((((1s,3s)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((cis-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((cis-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((cis-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((cis-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((cis-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((((1R,3S)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((((1s,3S)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((((1s,3S)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((((1s,3R)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((((1s,3s)-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((trans-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((trans-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((trans-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((trans-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((trans-3-methoxycyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((((1r,4S)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((((1r,4R)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((((1r,4R)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((((1r,4S)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((((1r,4r)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((4-methyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((4-methyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((4-methyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((4-methyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((4-methyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((pyrimidin-5-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((pyrimidin-5-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((pyrimidin-5-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((pyrimidin-5-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((pyrimidin-5-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((2-(tetrahydro-2H-pyran-4-yl)ethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((2-(tetrahydro-2H-pyran-4-yl)ethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((2-(tetrahydro-2H-pyran-4-yl)ethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((2-(tetrahydro-2H-pyran-4-yl)ethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((2-(tetrahydro-2H-pyran-4-yl)ethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(2-oxa-7-azaspiro[3.5]nonan-7-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((4-methoxytetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((4-methoxytetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((4-methoxytetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((4-methoxytetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((4-methoxytetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((7-oxaspiro[3.5]nonan-2-yl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((7-oxaspiro[3.5]nonan-2-yl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((7-oxaspiro[3.5]nonan-2-yl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((7-oxaspiro[3.5]nonan-2-yl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((7-oxaspiro[3.5]nonan-2-yl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 1-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)cyclobutane-1-carbonitrile; 1-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)cyclobutane-1-carbonitrile; 1-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)cyclobutane-1-carbonitrile; 1-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)cyclobutane-1-carbonitrile; 1-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)cyclobutane-1-carbonitrile; 3-(5-(((1S,2S)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(2-chlorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(2-methoxyphenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-((pyrazolo[1,5-a]pyrimidin-6-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-((pyrazolo[1,5-a]pyrimidin-6-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-((pyrazolo[1,5-a]pyrimidin-6-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-((pyrazolo[1,5-a]pyrimidin-6-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-((pyrazolo[1,5-a]pyrimidin-6-ylmethyl)amino)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((4,4-difluorocyclohexyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((4,4-difluorocyclohexyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((4,4-difluorocyclohexyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((4,4-difluorocyclohexyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((4,4-difluorocyclohexyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((2,4-difluorobenzyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((2,4-difluorobenzyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((2,4-difluorobenzyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((2,4-difluorobenzyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((2,4-difluorobenzyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((1R,2R)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((1R,2R)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((1R,2R)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((1R,2R)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((1R,2R)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((1S,2S)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((1S,2S)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((1S,2S)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((1S,2S)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((1S,2S)-2-methoxycyclopentyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)bicyclo[1.1.1]pentane-1-carbonitrile; 3-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)bicyclo[1.1.1]pentane-1-carbonitrile; 3-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)bicyclo[1.1.1]pentane-1-carbonitrile; 3-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)bicyclo[1.1.1]pentane-1-carbonitrile; 3-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)bicyclo[1.1.1]pentane-1-carbonitrile; 3-(5-(((1S,2S)-2-(3-(3-fluorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-(3-fluorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-(3-fluorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-(3-fluorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-(3-fluorophenoxy)azetidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3,3-difluorocyclobutyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((((1S,4R)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((((1s,4S)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((((1s,4S)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((((1s,4R)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((((1s,4s)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((cis-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((trans-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(bis(((1R,4S)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(bis(((1r,4R)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(bis(((1r,4R)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(bis(((1r,4S)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(bis(((1r,4R)-4-methoxycyclohexyl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)amino)methyl)benzonitrile; 4-((((1S,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)(methyl)amino)methyl)benzonitrile; 4-((((1R,2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)(methyl)amino)methyl)benzonitrile; 4-((((1R,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)(methyl)amino)methyl)benzonitrile; 4-((((1S,2R)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)(methyl)amino)methyl)benzonitrile; 4-(((2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)cyclohexyl)(methyl)amino)methyl)benzonitrile; 3-(5-(((1S,2S)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3,3-difluoropyrrolidin-1-yl)cyclohexyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(ethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(ethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(ethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(ethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(ethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(benzylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(benzylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(benzylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(benzylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(benzylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(diethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(diethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(diethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(diethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(diethylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(4-methoxy-4-methylpiperidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(4-methoxy-4-methylpiperidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(4-methoxy-4-methylpiperidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(4-methoxy-4-methylpiperidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(isobutylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(isobutylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(isobutylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(isobutylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(isobutylamino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(propylamino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(propylamino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(propylamino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(propylamino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(propylamino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(2-oxa-6-azaspiro[3.3]heptan-6-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-((((1R,4S)-4-methoxycyclohexyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-((((1r,4R)-4-methoxycyclohexyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-((((1r,4R)-4-methoxycyclohexyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-((((1r,4S)-4-methoxycyclohexyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-((((1r,4r)-4-methoxycyclohexyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cycloheptyl)oxy) isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(((tetrahydro-2H-pyran-4-yl)methyl)amino)cycloheptyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((3-methyloxetan-3-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3-methyloxetan-3-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3-methyloxetan-3-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3-methyloxetan-3-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3-methyloxetan-3-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-methoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-methoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-methoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-methoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-methoxyazetidin-1-yl)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(((3,3-difluorocyclobutyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(((3,3-difluorocyclobutyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(((3,3-difluorocyclobutyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(((3,3-difluorocyclobutyl)methyl)amino)cycloheptyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(diethylamino)-3-methylcyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(3-(pyridazin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(3-(pyridazin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(3-(pyridazin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(3-(pyridazin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(3-(pyridazin-3-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(3-(2,2,2-trifluoroethoxy)azetidin-1-yl)cyclopentyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3,3-dimethylpiperidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(3-isopropoxyazetidin-1-yl)cyclopentyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S,3S,4R)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S,3R,4R)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R,3R,4R)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R,3S,4R)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,4R)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S,3S,4S)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S,3R,4S)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R,3R,4S)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R,3S,4S)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,4S)-3-(3-ethoxyazetidin-1-yl)bicyclo[2.2.1]heptan-2-yl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2S)-2-(3-(pyrazin-2-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1S,2R)-2-(3-(pyrazin-2-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2R)-2-(3-(pyrazin-2-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-(((1R,2S)-2-(3-(pyrazin-2-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(1-oxo-5-((2-(3-(pyrazin-2-yloxy)azetidin-1-yl)cyclohexyl)oxy)isoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-aminocyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-aminocyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-aminocyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-aminocyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(diethylamino)cyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(diethylamino)cyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(diethylamino)cyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(diethylamino)cyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-(diethylamino)cyclopentyl)oxy)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-aminocyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-aminocyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-aminocyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-aminocyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-((2-aminocyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2S)-2-(diethylamino)cyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1S,2R)-2-(diethylamino)cyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2R)-2-(diethylamino)cyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; 3-(5-(((1R,2S)-2-(diethylamino)cyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; and 3-(5-((2-(diethylamino)cyclopentyl)oxy)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione; or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.
 15. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.
 16. The pharmaceutical composition of claim 15 further comprising at least one additional pharmaceutical agent. 